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"Penelitian dilakukan untuk mengetahui pengaruh kombinasi gliserol 6% dengan beberapa konsentrasi kuning telur terhadap kualitas spermatozoa ikan kerapu kertang Epinephelus lanceolatus (Bloch, 1970) pascakriopreservasi. Semen ikan kerapu kertang didapatkan dengan metode pengurutan. Larutan pengencer terdiri atas marine fish ringer, gliserol 6%, dan berbagai konsentrasi kuning telur (0%; 5%; 10%; 15%; 20%; dan 25%). Ekuilibrasi dilakukan selama 10 menit pada suhu 4 ºC. Pembekuan dalam freezer (-20 ºC) selama 48 jam. Pencairan pada suhu 45 ºC selama 30-60 detik. Evaluasi semen dilakukan secara makroskopis (warna, volume, dan pH) dan secara mikroskopis (motilitas, viabilitas, dan abnormalitas) serta kemampuan fertilisasinya terhadap telur ikan kerapu macan. Berdasarkan hasil uji ANAVA yang dilanjutkan dengan uji Tukey, terdapat perbedaan nyata (P>0,05) terhadap motilitas, viabilitas, dan kemampuan fertilisasi spermatozoa ikan kerapu kertang pascakriopreservasi, akan tetapi tidak berpengaruh nyata terhadap abnormalitas spermatozoa pascakriopreservasi (P>0,05). Hasil penelitian menunjukkan bahwa perlakuan kuning telur 15% merupakan konsentrasi optimum karena menghasilkan nilai persentase rata-rata motilitas, viabilitas, dan kemampuan fertilisasi tertinggi masing-masing sebesar 83,64 ± 1,72%, 81,44 ± 2,06%, dan 77,31 ± 1,90%. Nilai rata-rata terendah pada persentase abnormalitas sebesar 21,50 ± 1,20%.

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Research on the effect of combination 6% glycerol and eggyolk as cryoprotectant of giant grouper Epinephelus lanceolatus (Bloch, 1970) spermatozoa quality postcryopreservation. Giant grouper cement is obtained by sorting method. The diluent solution used consist of marine fish ringer, 6% glycerol, and varian egg yolk concentrations (0%; 5%; 10%; 15%; 20%; dan 25%). Equilibration is carried out for 10 minutes at 4ºC. Freezing in the freezer (-20 ºC) for 48 hours. Thawing at 45 ºC for 30-60 seconds. Cement evaluation is carried out macroscopically (color, volume, and pH) and microscopically (motility, viability, and abnormality) and also the ability to fertilize the egg of tigger grouper. Based on the ANAVA statistical test followed by Tukey, there were significant differences effect on motility, viability, and fertilization ability of spermatozoa of giant grouper postcryopreservation (P> 0.05), but abnormality was significantly affected (P <0.05). (P> 0.05). The optimum concentration of egg yolk is 15% because it produces the highest percentage value of motility, viability, and fertilization ability of 83.64 ± 1.72%, 81.44 ± 2.06% and 77.31 ± 1.90% respectively. The lowest average value of abnormalitiy percentage is 21.50 ± 1.20%."

"ABSTRAKKasus infeksi measles baik individu maupun kejadian luar biasa KLB di Indonesia masih banyak ditemui. Konfirmasi infeksi measles klinis hanya dapat dilakukan di Laboratorium Nasional. Keterbatasan kit komersial yang rutin digunakan menyebabkan pemeriksaan menjadi terhambat. Pengembangan in house plate coating spesifik IgM measles dengan indirect ELISA dilakukan dengan memodifikasi dan mengoptimasi microtiter plate dengan kultur virus measles. Kultur virus measles didapat dengan menumbuhkannya pada kultur sel vero/hSLAM. Optimasi plate coating dilakukan menggunakan kultur virus measles dengan isolat MO/38/V/07 dan J/10/358/Riau dalam pengenceran 1:1 mdash;1:2.048 dan inaktif atau tidaknya virus pada saat coating dilakukan. Optimasi pemeriksaan indirect ELISA untuk in house plate coating dilakukan dengan konsentrasi konjugat 1:10, 1:25, dan 1:50. In house plate coating telah dioptimasi dan menunjukkan hasil optimum untuk mendeteksi IgM measles pada pengenceran 1:16 dengan isolat MO/38/V/07 dalam keadaan inaktif dan pemeriksaan menggunakan konsentrasi konjugat 1:25.

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ABSTRAKCases of infection measles both individuals and extraordinary events outbreak in Indonesia are still widely encountered. Confirmation of measles clinical infection can only be done at the National Laboratory. The limitations of commercial kits that are routinely used cause the examination to be inhibited. The development of a specific IgM measles in house plate coating with indirect ELISA is done by modifying and optimizing the microtiter plate with measles virus culture. Viral culture measles obtained by growing it on cell culture vero hSLAM. Optimization of plate coating was done using culture of measles virus with MO 38 V 07 and J 10 358 Riau isolates in dilution of 1 1 mdash 1.2048 and inactivation or active of virus at the time of coating. Optimization of indirect ELISA examination for in house plate coating is done with conjugate concentration 1 10, 1 25, and 1 50. In house plate coating was optimized and showed optimum results for detecting IgM measles at 1 16 dilution with MO 38 V 07 isolates in inactivation and examination indirect ELISA using a 1 25 conjugate concentration."

"Enzim merupakan biokatalisator yang banyak digunakan di bidang industri, terutama deterjen, farmasi, makanan bahkan pemurnian minyak. Salah satu enzim yang banyak digunakan untuk pemurnian minyak ialah lysophospholipase. Sebanyak 50 kebutuhan enzim industri diperoleh dari mikroorganisme. Akan tetapi umumnya produk aktivitas enzim oleh mikroba galur liar kurang memadai untuk aplikasi di industri, sehingga perlu dilakukan rekayasa genetik. Pengklonaan gen penyandi lysophospholipase pernah dilakukan di Aspergillus niger dan Cryptococcus neoformans, tetapi belum pernah dilakukan dari bakteri alkalotermofilik. Bacillus halodurans CM1 merupakan bakteri alkalotrmofilik isolat BPPT. Penelitian terdahulu menujukkan bahwa bakteri tersebut memiliki enzim lipase, tetapi belum diteliti lebih lanjut mengenai jenis dan lipase rekombinannya. Penelitian ini bertujuan untuk mengklona gen penyandi lysophospholipase dari Bacillus halodurans CM1 ke Escherichia coli DH5? menggunakan vektor pGEM-T easy. Plasmid rekombinan tersebut disekuensing. Hasil penelitian diperoleh fragmen gen penyandi lysophospholipase yang berukuran 783 pasang basa serta tingkat homologi 100 dengan genom Bacillus halodurans C-125 yang menyandi gen lysophospholipase No akses GenBank: BA000004.3.

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Enzyme is a biocatalyst widely used in industry, for example detergent, pharmaceutical, food or oil purification. One of the most widely used enzymes for oil purification is lysophospholipase. As much as 50 of industrial enzyme needs are obtained from microorganisms. However, enzyme productivty from wild type microbial strain is usually limited and not applicable in industry, so that genetic engineering is necessary. Cloning gene encoding for lysophospholipase was once performed in Aspergillus niger and Cryptococcus neoformans, but has never been done from alkalothermophilic bacteria, such as Bacillus halodurans. Bacillus halodurans CM1 is an isolate of BPPT. Previous research has shown that this bacteria have lipase enzymes, but the study about their propertieshave not been conducted. This study aims to clone the gene t lysophospholipase from Bacillus halodurans CM1 to Escherichia coli DH5 using the pGEM T easy vector. The recombinant plasmid is sequenced. The results is gene fragment encoding lysophospholipase obtained with size 783 base pairs and 100 similiraty with gene encoding lysophospholipase from Bacillus halodurans C 125 No access GenBank BA000004.3."

"ABSTRAKGallus gallus domesticus atau yang dikenal sebagai ayam kampung merupakan hasil domestikasi dari jenis ayam hutan merah Gallus gallus gallus. Kabupaten Bangkalan merupakan salah satu Kabupaten yang terletak di Pulau Madura yang memiliki peternakan ayam ketawa hasil domestifikasi dari Kabupaten Sidrap. Penelitian ini bertujuan untuk mengetahui keragaman ayam ketawa dan solusi pemuliaan ayam ketawa untuk mempertahankan sumber daya hayati, oleh karena itu dilakukan penelitian ayam ketawa berdasarkan analisis bioakustik, morfometrik, dan DNA barcoding. Sampel ayam ketawa yang berasal dari Kecamatan Kamal mempunyai rata-rata durasi kokok terpanjang yaitu 6,00 3,0 detik dan rata-rata jumlah suku kata terbanyak berasal dari ayam ketawa Kecamatan Socah yaitu 7,20 5,80. Hasil analisis morfometrik seluruh populasi ayam ketawa di Kabupaten Bangkalan terdapat delapan karakter morfologi yang berbeda signifikan antara populasi ayam ketawa di Kecamatan Kamal dan Kecamatan Burneh yaitu bobot badan, panjang femur, panjang shank, lingkar shank, panjang sayap, tinggi jengger, panjang jari ketiga, dan panjang tulang dada, sedangkan empat karakter morfologi berbeda signifikan antara populasi ayam ketawa di Kecamatan kamal dan Kecamatan Socah yaitu panjang shank, panjang sayap, panjang jari ketiga, dan panjang tulang dada, serta tiga karakter morfologi berbeda signifikan antar individu di Kecamatan Burneh dan Kecamatan Socah yaitu bobot badan, panjang femur, dan lingkar shank. Adanya perbedaan nilai signifikan karakter morfologi antar populasi disebabkan oleh faktor eksternal. Hasil nilai bootstrap tinggi yaitu 1000, sedangkan pada nilai bootstrap terkecil yaitu 382. Jarak genetik sebagai dasar rekonstruksi pohon filogeni yang dihasilkan dari populasi ayam ketawa di Kabupaten Bangkalan yaitu berkisar antara 0,025--1,872. Analisis molekuler melalui teknik DNA Barcoding dengan Gen Cytochrome Oxidase Sub Unit 1 COI dapat digunakan untuk mengidentifikasi spesies ayam ketawa di Kabupaten Bangkalan G. g. domesticus dan persebarannya.

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ABSTRACTGallus gallus domesticus or known as native chicken became domesticated from wild junglefowls Gallus gallus gallus. Bangkalan District is one of the districts located in Madura Island which has breeding domesticated chicken from Sidrap District. The purpose of this research is to know the diversity of ketawa chicken and ketawa chicken Breeding solution to preserve the biological resources, therefore the research of ketawa chicken ketawa is done based on bioacoustic, morphometric, and DNA barcoding analysis. Ketawa chicken samples from Kamal Subdistrict had the longest duration of crowing length of 6.00 3.0 seconds and the average number of syllables ketawa chicken was mostly from Socah Subdistrict of 7.20 5.80. Result of morphometric analysis of whole population of ketawa chicken in Bangkalan District there are eight morphological characters significantly different between ketawa chicken population in Kamal Subdistrict and Burneh Subdistrict are body weight, femur length, shank length, shank circumference, wing length, height of comb, third finger length, and chest length, where as four morphological characters were significantly different between ketawa chicken population in Kamal Subdistrict and Socah Subdistrict, are shank length, wing length, finger length, and chest length, and three significantly different morphological characters between individuals in Burneh Subdistrict and Socah Subdistrict namely body weight, femur length, and shank circumference. The existence of significant difference of morphological character between population is caused by external factor. The result of high bootstrap value is 1000, while at the smallest bootstrap value is 382. Genetic distance as the basis of phylogenetic tree reconstruction resulting from ketawa chicken population in Bangkalan District is ranged from 0,025 1,872. Molecular analysis through DNA Barcoding technique with Cytochrome Oxidase Sub Unit 1 COI Genes can be used to identify Ketawa chicken species in Bangkalan District G. g. domesticus and their distribution."

"Ikan mas mustika merupakan strain unggulan dan merupakan komoditi bernilai ekonomi tinggi di Jawa Barat. Peningkatan jumlah ketersediaannya, memerlukan induk yang berkualitas. Pemijahan ikan dipengaruhi oleh cuaca. Alternatif perlu ditemukan untuk percepatan pematangan gonad pada ikan mas mustika, untuk membantu dalam ketersediaanya. Bahan umum yang digunakan dalam budidaya ikan mas mustika adalah SnGrH dengan merek dagang ovaprim. Penggunaan hormon sintetik dinilai tidak ekonomis dan bisa berdampak buruk bagi ikan jika dilakukan terus-menerus. Bahan alternatif yang ekonomis dan aman berasal tanaman. Cabe jawa merupakan salah satu tanaman afrodisiak yang memiliki efek hormonal, serta memiliki pengaruh terhadap reproduksi yaitu piperin dan sitosterol. Penelitian ini bertujuan untuk mengevaluasi pengaruh konsentrasi ekstrak cabe jawa terhadap kualitas sperma ikan mas mustika. Penelitian ini dilakukan dengan menggunakan ikan mas mustika yang berumur tiga bulan. Terdapat tiga jenis perlakuan pemberian ekstrak cabe jawa yaitu (ECJ) 7,8 mL mg-1 kg-1 bobot ikan, 15,6 mL mg-1 kg-1 bobot ikan dan 31,2 mL mg-1 kg-1 bobot ikan. NaCl diberikan sebagai kontrol negatif dan ovaprim sebagai kontrol positif. Penyuntikan dilakukan sekali yaitu di minggu pertama dan pengambilan sampel dilakukan pada minggu kedua setelah penyuntikan. Sampling sperma dilakukan dengan cara stripping. Sperma dievaluasi secara makroskopis (volume, pH, dan warna), mikroskopis (konsentrasi sperma, motilitas dan durasi motilitas), dan hasil dari sperma juga digunakan untuk fertilisasi, yang dilakukan secara buatan. Hasil penelitian menunjukkan bahwa penggunaan ekstrak cabe jawa dengan konsentrasi 7,8 mL mg-1 kg-1 bobot ikan (perlakuan A) berpengaruh nyata (P < 0,05) terhadap konsentrasi sperma, motilitas, durasi motilitas dan fertilisasi. Berdasarkan hasil tersebut dapat disimpulkan bahwa pemberian ekstrak cabe jawa terhadap kualitas sperma mempengaruhi dan meningkatkan kualitas sperma serta fertilisasi. Hasil optimum yaitu pada konsentrasi 7,8 mL mg-1 kg-1 ikan dengan hasil konsentrasi sperma (58,67 ±1,45 )105 sel/mL dengan nilai (69,5± 1,24)%, durasi motilitas (32,5 ±1,7) detik, dan fertilisasi (66,50 ± 1,27)%. Mustika common carp is one of strain with good quality than the other carp and being popular quality in West Java. Increase production requires the restock of fish with high quality. Procurement of fish seeds is affected by weather, need to accelerate gonadal maturation in common carp mustika. Common materials used in aquaculture for common carp mustika is SnGrH with brand name ovaprim. The synthetic hormone is considered uneconomical and have effects which is not safe for fish. It is necessary to use economical and safe alternative ingredients such as java long pepper. Java long paper is one of the aphrodisiac plants that have hormonal efficiency. The active ingredient is thought to have an effect on reproduction, namely piperine and sitosterol. This study aims to evaluate the effect of long pepper extract for quality of common carp mustika spermatozoa. This study was conducted using 3 months old common carp mustika fishes. Treatments including giving three concentration of java long pepper extract were 7.8 mL mg-1 kg-1 weight body, 15.6 mL mg-1 kg-1 weight body and 31.2 mL mg-1 kg-1 weight body. The negative control used NaCl physiological solution and also positive control by SnGrH (ovaprim). Injection was given on first week, and did frequently checked everyday and at second week fish can product spermatozoa. Spermatozoa was evaluated by microscopic factors (volume, pH, and colors), by macroscopic factors (Sperm density, motility and motility duration) also fertilization. Result showed that using extract long paper with doses 7.8 mL mg-1 kg-1 weight body have a real impact (P <0.05) on sperm density, motility and motility duration. Highest result for sperm concentration (58.67± 1.45)105 sel/mL, motility (69,5±1,24)%, and motility duration (32.5 ±1,7)seconds. "

"Nerve Growth Factor (NGF) diketahui memiliki konsentrasi yang cenderung lebih tinggi pada wanita endometriosis dibandingkan dengan wanita tanpa endometriosis. Metilasi DNA pada daerah promotor NGF diyakini sebagai salah satu penyebab meningkatnya konsentrasi NGF pada wanita endometriosis. Penelitian ini dilakukan untuk mengetahui korelasi antara metilasi DNA dengan ekspresi mRNA NGF pada darah menstruasi wanita endometriosis. Sebanyak 10 sampel darah menstruasi dari masing-masing wanita endometriosis dan wanita tanpa endometriosis digunakan dalam penelitian ini. Tingkat metilasi DNA NGF dihitung menggunakan Methylation-specific Polymerase Chain Reaction (MSP-PCR), kemudian intensitas pita yang muncul diukur menggunakan software ImageJ. Nilai ekspresi mRNA NGF dihitung menggunakan Quantitative Real-Time Polymerase Chain Reaction (qRT PCR). Analisis korelasi, selanjutnya, dilakukan menggunakan aplikasi SPSS. Hasil penelitian menunjukkan bahwa tingkat metilasi DNA NGF memiliki tendensi yang rendah pada darah menstruasi wanita endometriosis sebesar 35,27%. Ekspresi relatif mRNA NGF menunjukkan tendensi peningkatan 19,229 lebih tinggi pada darah menstruasi wanita endometriosis. Tingkat metilasi DNA dan ekspresi mRNA NGF tidak menunjukkan perbedaan baik pada darah menstruasi wanita endometriosis maupun wanita tanpa endometriosis dengan nilai p > 0,05. Korelasi metilasi DNA dengan ekspresi mRNA NGF pada darah menstruasi wanita endometriosis tidak dapat dianalisis karena data yang tersedia sangat terbatas.

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Nerve Growth Factor (NGF) is known to have higher concentrations in women with endometriosis compared to women without endometriosis. DNA methylation in the NGF promoter region is believed to cause the increase of NGF concentrations in women with endometriosis. This study is conducted to determine the correlation between DNA methylation and mRNA expression of NGF in menstrual blood of women with endometriosis. A total of 10 menstrual blood samples from women with endometriosis and women without endometriosis were used in this study. DNA methylation level of NGF was measured using Methylation-specific Polymerase Chain Reaction (MSP-PCR) and the intensity of the bands were subsequently measured using ImageJ software, while mRNA expression values of NGF ​​were measured using Quantitative Real-Time Polymerase Chain Reaction (qRT PCR). Furthermore, the correlation analysis was performed using the SPSS application. The results exhibited that the methylation level of NGF shows a low tendency in menstrual blood of women with endometriosis of 35.27%. The relative mRNA expression of NGF showed a tendency to increase 19,229-fold higher in menstrual blood of women with endometriosis. The DNA methylation level and mRNA expression of NGF showed no difference in both menstrual blood of women with endometriosis and women without endometriosis with p value > 0,05. The correlation between DNA methylation and mRNA expression of NGF in menstrual blood of women with endometriosis could not be analyzed because of very limited data."

"Efisiensi transformasi plasmid rekombinanyang rendah ke dalam bakteri wild-type disebabkan oleh mekanisme pertahanan dari sel bakteri. Enzim restriksi dalam sel bakteri target dapat mendegradasi plasmid rekombinan. Transformasi plasmid pBBRE194 rekombinan yang mengandung gen protease dari Bacillus halodurans CM1 (pBBRE194 prot-CM1) telah dilakukan, tetapi efisiensi transformasi rendah dan klona rekombinan yang didapatkan tidak menunjukkan sifat yang stabil. Penelitian ini bertujuan untuk memetilasi plasmid pBBRE194 prot-CM1 dan transformasi plasmid pBBRE194 prot-CM1 termetilasi secara konjugasi ke B. halodurans CM1, kemudian menganalisis aktivitas enzim protease yang dihasilkan B. halodurans CM1 pembawa plasmid pBBRE194 prot-CM1. Aktivitas spesifik (U/mg) protease yang dihasilkan oleh B. halodurans CM1 rekombinan dianalisis dengan cara mengukur unit aktivitas (U/mL) dengan metode Amano dan kadar protein (mg/mL) dengan metode Bradford. Plasmid pBBRE194 prot-CM1 dalam E. coli TOP10 berhasil dimetilasi oleh gen methylase pada plasmid pPAMC125 yang diinduksi oleh 0,02% L-arabinosa. Konjugasi plasmid pBBRE194 prot-CM1 termetilasi ke B. halodurans CM1 berhasil dilakukan dan terpilih 1 klona B. halodurans CM1 rekombinan yang telah terverifikasi. Verifikasi dilakukan berdasarkan kemampuan klona dalam mendegradasi protein pada media selektif yang mengandung skim milk dan tetracycline. Verifikasi berdasarkan polymerase chain reaction dengan mendeteksi sekuens gen resistan tetracycline pada klona juga dilakukan. Proses PCR koloni pada B. halodurans CM1 rekombinan menunjukkan terbentuknya pita DNA ukuran 1024 bp yaitu ukuran gen resistan tetracycline pada plasmid pBBRE194 prot-CM1. Hasil analisis menunjukkan bahwa aktivitas spesifik B. halodurans CM1 rekombinan (660,700 U/mg) lebih rendah dibandingkan dengan kontrol negatif, yaitu B. halodurans CM1 wild-type (1054,928 U/mg). Analisis aktivitas protease dilakukan pada suhu 50 oC dan pH 12.

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Transformation rate into wild-type bacteria is commonly low because of the cell defense mechanism of the bacteria. Restriction modification (RM) in bacteria cells can prevent the introduction of recombinant plasmid into target bacteria. Previously, the transformation of recombinant shuttle vector pBBRE194 containing protease gene (pBBRE194 prot-CM1) into wild-type Bacillus halodurans CM1 has been conducted. However, the transformation rate seemed low, and the stable recombinant clones could not be obtained. Therefore, in vivo methylation of this plasmid in E. coli has to be done before genetic transformation into the wild-type bacterium, to obtain stable recombinant B. halodurans CM1. In this study, a plasmid with artificial modification (pPAMC125) harboring genes encoding for the modification enzymes (methylases) from another strain, B. halodurans C-125, and pBBRE194 prot-CM1 plasmid were transformed by conjugation into B. halodurans CM1. Specific activity (U/mg) of protease produced by recombinant B. halodurans CM1 was analyzed by measuring activity units (U/mL) by the Amano method and protein quantity (mg/mL) by the Bradford method. The pBBRE194 prot-CM1 might be methylated by methylases that was induced by 0.02% L-arabinose. Conjugation of the methylated pBBRE194 prot-CM1 to B. halodurans CM1 was successfully carried out and recombinant B. halodurans CM1 was verified. The verification of a recombinant clone is based on its ability to degrade proteins on selective media containing skim milk and tetracycline. Also beside, verification based on polymerase chain reaction was also carried out by detecting tetracycline resistance gene sequences. The PCR result in recombinant clone amplified the 1024 bp DNA, which the size of the tetracycline resistance gene in pBBRE194 prot-CM1 plasmid. The analysis of protease activity showed that the specific activity of the recombinant clone (660.700 U/mg) was lower than the negative control, which B. halodurans CM1 wild-type (1054.928 U/mg). The analysis was carried out at 50oC and pH 12. "

"Bacillus halodurans CM1 berpotensi sebagai inang dalam menghasilkan beberapa jenis enzim yang berguna, seperti xilanase, lipase, dan protease. Selain sebagai penghasil enzim, salah satu gen potensial lainnya adalah gen resistan antibiotik. Penelitian ini bertujuan untuk mengetahui kemampuan resistansi B. halodurans CM1 terhadap eritromisin dan kanamisin serta diperoleh produk gen fungsional resistan eritromisin dan kanamisin dari B. halodurans CM1. Isolasi gen eritromisin dan kanamisin dari B. halodurans CM1 belum pernah dilakukan. Berdasarkan uji KHM, B. halodurans CM1 resistan terhadap eritromisin dan kanamisin. Isolasi gen resistan eritromisin dan kanamisin dilakukan dengan menggunakan metode amplifikasi PCR. Produk PCR yang diduga gen resistan eritromisin, yaitu gen ErmK dan BH0381. Produk PCR yang diduga gen resistan kanamisin, yaitu gen aadK dan APH. Gen ErmK dikloning dengan menggunakan kloning vektor pGEM-T Easy, sedangkan gen BH0381, aadK, dan APH dikloning dengan menggunakan kloning vektor pJET1.2/blunting. Vektor rekombinan ditransformasi ke Escherichia coli DH5alfa. Hasil analisis sekuens DNA menggunakan BLAST menunjukkan bahwa gen ErmKCM1 dan BH0381 masing-masing memiliki kemiripan 99,65% (GenBank No access: BH0380, ErmK) dan 99,45% (GenBank No access: BH0381, mphB) dengan sekuens dari B. halodurans C-125. Sekuens gen ErmKCM1 dan BH0381CM1 menunjukkan bahwa dua gen tersebut merupakan gen yang fungsional. Sekuens upstream dari gen BH0381CM1 dianalisis dan diperoleh bahwa terdapat 50 pb yang diduga promoter. Hasil analisis sekuens DNA menggunakan BLAST menunjukkan bahwa gen aadKCM1 dan APHCM1 masing-masing memiliki kemiripan 97,73% (GenBank No access: BH0322) dan 99,47% (GenBank No access: BH0326) dengan sekuens dari B. halodurans C-125. Hasil penyejajaran gen aadKCM1 menunjukkan adanya delesi enam pasang nukleotida pada lokus ke-354 hingga 359 yang menyebabkan frameshift, sehingga gen aadKCM1 tidak memiliki sekuens yang open reading frame (ORF). Hasil translasi gen aadKCM1 dan APHCM1 menunjukkan bahwa hanya sekuens gen APHCM1 dapat ditranslasi menjadi protein yang fungsional. Hasil uji resistansi kanamisin pada transforman yang membawa plasmid rekombinan promoter gen APHCM1-ORF dapat menyandikan resistan kanamisin dengan konsentrasi kanamisin sebesar 20 µg/mL.

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Bacillus halodurans CM1 has potential as a host in producing several types of useful enzymes, such as xylanase, lipase, and protease. Besides industrial enzymes, one of the other potential genes is the antibiotic-resistant gene. This study aims to determine the ability of B. halodurans CM1 resistance to erythromycin and kanamycin and to obtain erythromycin and kanamycin-resistant functional gene products from B. halodurans CM1. Isolation of erythromycin and kanamycin genes from B. halodurans CM1 has never been done. Based on the MIC test, B. halodurans CM1 is resistant to erythromycin and kanamycin. Erythromycin and kanamycin resistance gene isolation was carried out using PCR amplification method. PCR products suspected of being erythromycin resistance genes, namely ErmK and BH0381 genes. PCR products suspected of being kanamycin resistance genes, namely genes aadK and APH. The ErmK gene was cloned using the pGEM-T Easy vector cloning, while the BH0381, aadK, and APH genes were cloned using the pJET1.2/blunting vector cloning. The recombinant vector was transformed into Escherichia coli DH5ï?¡. The results of DNA sequence analysis using BLAST showed that the ErmKCM1 and BH0381 genes each had 99.65% similarities (GenBank No access: BH0380, ErmK) and 99.45% (GenBank No access: BH0381, mphB) with the sequence of B. halodurans C-125. The results of the translation of the ErmKCM1 and BH0381CM1 genes indicate that the two genes are functional genes. The upstream sequence of the BH0381CM1 gene was analyzed and it was found that 50 bp was suspected as a promoter. The results of DNA sequence analysis using BLAST showed that the aadKCM1 and APHCM1 genes each had a similarity of 97.73% (GenBank No access: BH0322) and 99.47% (GenBank No access: BH0326) with the sequence of B. halodurans C-125. The alignment of the aadKCM1 gene shows the deletion of six nucleotide pairs at the 354-359 locus that causes frameshift, so the aadKCM1 gene does not have an open reading frame (ORF) sequence. The translation of aadKCM1 and APHCM1 genes show that only the APHCM1 gene sequence can be translated into a functional protein. The results of kanamycin resistance test on transformants carrying plasmids with native promoter APHCM1-ORF gene can encode kanamycin resistance with a kanamycin concentration of 20 µg/mL."