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"Latar belakang: Antigen38 Mycobacterium tuberculosis merupakan agen serodiagnostik yang potensial karena mengandung
dua epitop spesifik untuk sel B. Mahalnya agen diagnostik menyebabkan lambatnya realisasi diagnosis TB secara cepat di
negara berkembang. Kami memproduksi antigen 38 rekombinan yang berasal dari galur lokal yang kemungkinan dapat
digunakan untuk memproduksi alat serodiagnostik TB yang ekonomis.
Metode: Gen pab diisolasi dari pasien TB paru di Malang, diklon ke plasmid pGEM-Teasy menjadi pMB38. Klon E.coli
DH5α yang membawa pMB38 diseleksi di medium yang ditambah dengan X-Gal. Ekspresi pab dilakukan menggunakan
pMBhis yang berasal dari pPRoExHTc dibawah kontrol promoter Trc dengan inang E.coli DH5α
Hasil: Pencocokan sekuen gen pab dari klon E.coli DH5α berwarna putih dengan gen pab dari M. tuberculosisH37Rv
memperlihatkan homologi sebesar 98%. Protein rekombinan yang sudah dihilangkan signal peptidanya ditemukan di
sitoplasma.
Kesimpulan: Gen pab dari pasien TB dapat diekspresikan secara intraseluler dengan sistem heterolog

Abstract
Background: Mycobacterium tuberculosis antigen38 is a potent serodiagnostic agent containing two M. tuberculosisspecific
B-cell epitopes. The high price of imported diagnostic agents hinders realization of fast clinical TB diagnosis in
developing countries. Therefore, we produced recombinant antigen38 (recAg38M) from M. tuberculosis local strain, which
might be used to produce economical tuberculosis serodiagnostic kit.
Methods: Pab gene that was isolated from pulmonary TB patient in Malang was cloned into a plasmid vector (pGEMTeasy)
to construct pMB38. The E.coli DH5α clone carrying pMb38 was selected on X-gal medium. The expression of pab
was mediated using pPRoExHTc under the control of Trc promoter and E.coli DH5α as host.
Results: Alignment of the pab sequence from the white E.coli DH5α clones with that of M. tuberculosis H37Rv showed
98% homology. The recombinant protein in which the signal peptide has been deleted to prevent the protein being secreted
into medium was found in the cytoplasm.
Conclusion: pab gene of M. tuberculosis isolated from a TB patient could be expressed in heterologous system in E.
coliDH5α."
[Fakultas Kedokteran Universitas Indonesia, Universitas Brawijaya. Fakultas Matematika dan Ilmu Pengetahuan Alam], 2011
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Artikel Jurnal  Universitas Indonesia Library
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"Untuk memperoleh koleksi kekayaan hayati Nostoc Indonesia dilakukan isolasi mikroflora tanah dari beberapa lahan persawahan di wilayah di Jawa, Bali, dan Sulawesi Selatan. Isolat-isolat yang tumbuh cepat diidentifikasi secara morfologi dan molekuler mengunakan sekuen parsial dari gen 16S rRNA. Walaupun hubungan kekerabatan antar isolat belum sepenuhnya dapat dijelaskan, pohon filogeni yang dihasilkan dari analisis sekuen mendukung identifikasi secara morfologi bahwa isolat-isolat yang diteliti berbeda jenis. Uji coba 6 strain Nostoc , dalam bentuk inokulum tunggal, sebagai sumber nitrogen untuk padi dilakukan. Sebanyak 2 g biomasa basah dari masing-masing strain Nostoc diinokulasi ke dalam pot-pot yang telah berisi 3 tanaman padi. Percobaan dilakukan di rumah kaca selama 115 hari. Secara statistik (ANOVA;α= 0.05) hanya strain GIA13a yang mempengaruhi panjang akar dan jumlah bulir padi bernas.

In order to collect Indonesian Nostoc, isolation of soil microflora from several paddy fields in West Java, Bali, and South Celebes was carried out. Fast-growing isolates of Nostoc were selected to describe and perform molecular identification using partial sequences of 16S rRNA. The results showed that partial sequences of 16S rRNA could not resolve the phylogeny of the isolates. However, it supported the morphological studies that recognize isolates as different species of Nostoc. Potential use of Nostoc as a nitrogen source for paddy growth was carried out using six strains as single inoculums. A total biomass of 2 g (fresh weight) for each strain was inoculated, respectively, into the pot planted with three paddy plants. This experiment was conducted in the green house for 115 days. Statistical analyses (ANOVA;α= 0.05) showed that of six strains tested in this study, only strain GIA13a had influence on the augmentation of root length and the total number of filled grains.
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Depok: Direktorat Riset dan Pengabdian Masyarakat Universitas Indonesia, 2012
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Artikel Jurnal  Universitas Indonesia Library
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Ira Trisnawati
"Asam lemak omega-3 merupakan asam lemak essensial bagi tubuh manusia. Mikroalga merupakan salah satu sumber asam lemak omega-3 yang sangat prospektif untuk dikembangkan, karena kualitas yang lebih tinggi dibanding sumber asam lemak omega-3 lainnya. Pemanfaatan langsung asam lemak omega-3 dari mikroalga memerlukan biaya produksi lebih tinggi. Sehingga penelitian terkini banyak diarahkan pada studi genetika terhadap enzim yang berperan penting dalam biosintesis asam lemak omega-3, salah satunya adalah enzim omega-3 desaturase. Penelitian ini akan difokuskan pada isolasi dan kloning gen yang mengkode enzim omega-3 desaturase dari mikroalga Nannochloropsis sp. Gradient Polymerase Chain Reaction ( PCR ) berhasil mengamplifikasi gen omega-3 desaturase dengan panjang 489bp. Gen disisipkan pada plasmid T-vector pMD20 dan dikloning pada sel kompeten bakteri Escherichia coli DH5α. Konfirmasi produk kloning melalui colony PCR dan dari hasil konfirmasi berat molekul yang dianalisa menggunakan metode agarose electrophoresis menunjukkan terdapat 6 koloni yang positif mengandung gen omega-3 desaturase. Konfirmasi dengan DNA sequencing masih perlu dilakukan di masa yang akan datang.

Omega-3 fatty acids are essential fatty acids for the human body. Microalgae is one source of omega-3 fatty acids which is highly prospective for development, because it has higher quality than the other sources of omega-3 fatty acids. Direct utilization of omega-3 fatty acids from microalgae requires higher production cost. Therefore, many of the recently studies focus on genetic study for the enzymes which play important role in biosynthesis of omega-3 fatty acids, one of them is omega-3 desaturase enzyme. This research will be focused on the isolation and cloning of gene encoding omega-3 desaturase enzyme from microalgae Nannochloropsis sp. Gradient Polymerase Chain Reaction (PCR) successfully amplified 489bp of omega-3 desaturase gene. The gene was inserted into T-Vector pMD20 plasmid and cloned to Escherichia coli DH5α competent cell. Confirmation cloning product using colony PCR and from molecular weight analyzing by agarose electrophoresis method showed that there are 6 colonies positively content omega-3 desaturase gene. Confirmation by DNA sequencing still needs to be done in the future."
Depok: Fakultas Teknik Universitas Indonesia, 2012
S43332
UI - Skripsi Open  Universitas Indonesia Library
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Washila Nurlaila
"ABSTRACT
Isolasi dan penapisan kapang dan khamir dari lima jenis ragi tapai asal beberapa kota di Jawa Barat telah dilakukan. Berdasarkan hasil isolasi, didapatkan tiga belas isolat kapang dan tujuh isolat khamir. Penapisan kapang amilolitik dilakukan secara kualitatif menggunakan uji iodin. Uji kualitatif dilakukan dengan mengukur zona bening pada medium starch agar yang telah ditumbuhi kapang dan kemudian ditetesi iodin. Hasil uji menunjukkan isolat (ZC1, ZC2, ZGJ2) memiliki diameter zona bening sebesar (69,95 mm, 58,73 mm, 56,85 mm). Aktivitas amilase ketiga isolat kapang terpilih diukur menggunakan metode DNS (Dinitrosalicylic Acid). Hasil uji menunjukkan bahwa isolat ZGJ2 merupakan isolat kapang dengan aktivitas tertinggi (6,30 U/mL) sedangkan isolat kapang dengan aktivitas terendah (3,03 U/mL) dihasilkan oleh isolat ZC2. Penapisan khamir penghasil alkohol dilakukan berdasarkan pertumbuhan sel dan gas  yang terperangkap dalam tabung Durham, dalam medium PDB yang ditambah glukosa 5%, 10%, dan 15%. Ketiga isolat mampu tumbuh dengan baik pada medium dengan konsentrasi glukosa 15%. Namun pembentukan gas  hanya terjadi pada penambahan 10% glukosa oleh isolat YC1 (4+) dan YC3 (3+)  serta penambahan 5% glukosa oleh isolat YC2 (2+). Hasil pengamatan karakter makroskopis dan mikroskopis isolat ZC1 dan ZGJ2  diduga merupakan genus Rhizopus, sedangkan isolat ZC2 masuk ke dalam genus Mucor. Isolat khamir terpilih diduga termasuk ke dalam filum Ascomycota berdasarkan karakter morfologi dan fisiologi.

ABSTRACT
Isolation and screening of molds and yeasts from five types of ragi tapai from several cities in West Java had been done. Based on the results of isolation, thirteen mold isolates and seven yeast isolates were obtained. Screening of amylolytic mold was done by qualitative assay using iodine. Iodine assay was done by measuring clear zones on starch agar medium which had been grown with mold and then flooded with iodine. The results of iodine assay showed that three isolates (ZC1, ZC2, ZGJ3) formed clear zones diameter (69.95, 58.73, 56.85). Amylase activity of the three selected mold isolates were measured using the DNS (Dinitrosalicylic Acid) method. The results showed that ZGJ2 had highest activity (6.30 U / mL) meanwhile the mold isolate with the lowest activity (3.03 U / mL) was ZC2. Alcohol-producing yeasts were screened based on cell growth and  trapped in Durham tubes, in the medium of PDB added with glucose 5%, 10%, and 15%. The best three isolates were able to grow in a medium with 15% glucose concentration. However the formation of  only occurs in the addition of 10% glucose by YC1 (4+) and YC3 (3+) and the addition of 5%  glucose by YC2 (2+). Based on observation of the macroscopic and microscopic characters, ZC1 and ZGJ2 assumed belong to the Rhizopus genus, meanwhile ZC2 belongs to the Mucor genus.The selected yeasts are assumed to belong to the Ascomycota phylum based on morphological and physiological characters."
2019
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UI - Skripsi Membership  Universitas Indonesia Library
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Widamayanti
"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.

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."
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2020
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UI - Tesis Membership  Universitas Indonesia Library
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Labeda, David P.
New York : McGraw-Hill , 1990
576 LAB i
Buku Teks SO  Universitas Indonesia Library
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"Latar belakang: Infeksi saluran cerna dengan manifestasi klinis berupa diare merupakan penyakit infeksi dengan kesakitan dan kematian yang tinggi terutama di negara-negara yang sedang berkembang. Diare menimbulkan kematian terutama pada bayi baru lahir di bawah 1 tahun. Penanganan telah ditingkatkan secara terus menerus, namun demikian kemajuan dalam diagnosis maupun pengobatan tidak terjangkau oleh negara-negara yang sedang berkembang. Salah satu penyebab infeksi saluran cerna adalah bakteri. Oleh karenanya dengan mengetahui bakteri penyebab serta pola resistensi bakteri terhadap antibiotik dapat menunjang penatalaksanaan penyakit ini. Studi ini dilakukan untuk mengetahui berbagai jenis mikroba yang diisolasi dari saluran cerna serta pola resistensinya terhadap beberapa antibiotik.
Metode: Spesimen berupa tinja, usap dubur atau anus yang diterima oleh Laboratorium Mikrobiologi FKUI selama 2005- 2008. Isolasi, identifi kasi kepekaan dan uji antibiotik dikerjakan sesuai prosedur standar yang berlaku. Interpretasi hasil uji kepekaan menggunaan panduan NCCLS/CLSI. Data dianalisis menggunakan WHOnet versi 5.3.
Hasil: Diperoleh 28 isolat Escherichia coli patogen, 1 isolat Salmonella paratyphi A, dan 4 isolat ragi yang diisolasi dari tinja dan swab dubur penderita. Walaupun Escherichia coli patogen masih peka terhadap beberapa antibiotik, namun kepekaannya menurun terhadap amoxicillin, sulbenicillin, ticarcillin dan trimethoprim/rulfamethoxazole.
Kesimpulan: Escherichia coli patogen merupakan bakteri terbanyak yang berhasil diisolasi dari tinja/usap dubur. Bakteri ini telah menunjukkan penurunan kepekaan terhadap beberapa antibiotik yang sering digunakan untuk mengobati infeksi saluran cerna. (Med J Indones 2011; 20:105-8).

Abstract
Background: Digestive tract infection with clinical manifestation of diarrhea is an infectious disease that has the highest morbidity and mortality rate, especially in developing countries. Diarrhea causes mortality mostly in infants under one year old. Improvement in management is done continuously, but advances in diagnosis and therapy cannot be reached by developing countries. One of the etiological agents causing infection of digestive tract is bacteria. Therefore, knowledge of bacteria that cause gastrointestinal infection and their resistance patterns may support the management of this disease. The aim of this study was to examine microbes that were isolated from the digestive tract and their resistance patterns against antibiotics.
Methods: Samples (stool, rectal/anal swab) were collected from the Clinical Microbiology Laboratory, FKUI during 2005-2008. Isolation, identifi cation and sensitivity test were conducted according to standard laboratory procedures. Interpretation of sensitivity test was done according to NCCLS/CLSI guidance. Data was analyzed using WHOnet version 5.3.
Results: We found 28 isolates of pathogenic Escherichia coli, 1 isolate of S. paratyphi A and 4 isolates of yeasts. Pathogenic Escherichia coli were still sensitive against some antibiotics, but the sensitivity was reduced against amoxicillin, sulbenicillin, ticarcillin and trimethoprim/sulfamethoxazole.
Conclusion: The most predominant gastrointestinal tract infection causing microbes was pathogenic Escherichia coli. These bacteria showed decrease sensitivity against some antibiotics commonly used to treat patients with gastrointestinal tract infection. (Med J Indones 2011; 20:105-8)"
[Fakultas Kedokteran Universitas Indonesia, Fakultas Kedokteran Universitas Indonesia], 2011
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Artikel Jurnal  Universitas Indonesia Library
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Ricky Karta Atmadja
"Telah dilakukan penelitian yang bertujuan untuk mengetahui aktivitas antimikroba actinomycetes termofil hasil isolasi dari geiser di Cisolok, Jawa Barat. Delapan belas isolat yang memiliki morfologi menyerupai actinomycetes berhasil diisolasi dari serasah daun dan ranting di sekitar pusat semburan geiser. Seluruh isolat diuji aktivitas antimikrobanya menggunakan paper disk method dan agar block method dengan Kocuria rhizophila, Staphylococcus aureus, Bacillus subtilis sebagai bakteri uji Gram positif, dan Escherichia coli sebagai bakteri uji Gram negatif. Pengujian menggunakan metode paper disk menunjukkan hasil negatif pada isolat actinomycetes yang dikultur pada medium International Streptomyces Project (ISP) 1 cair selama 14 hari pada suhu 50oC dan 40oC. Berdasarkan uji menggunakan metode blok agar, didapatkan bahwa dua isolat, yaitu LC2-2 dan LC2-6 memberikan hasil positif terhadap bakteri uji Gram positif. Isolat LC2-2 menunjukkan morfologi makroskopis dan mikroskopis menyerupai genus Bacillus sehingga tidak digunakan untuk identifikasi molekuler. Hasil identifikasi molekuler sequence parsial gen 16S rRNA menggunakan primer 785F dan primer 802R menunjukkan bahwa LC2-6 diidentifikasi sebagai Actinomadura keratinilyitica dengan nilai homologi 99%. Berdasarkan hasil penelitian, direkomendasikan untuk mempelajari lebih lanjut senyawa antimikroba yang dihasilkan isolat LC2-6. Hal tersebut disebabkan oleh belum adanya laporan penelitian mengenai aktivitas antimikroba Actinomadura keratinilytica.

The aim of this study was to screen the antimicrobial activity by actinomycetes isolated from Cisolok Geyser, West Java. Eighteen isolates which are have similar morphology with actinomycetes have been isolated from leaves and branches around the geyser. The isolates were screened for their antimicrobial activity using paper disk method and agar block method with Kocuria rhizophila, Staphylococcus aureus, Bacillus subtilis as Gram positive test bacteria and Escherichia coli as Gram negative test bacteria. Screening by paper disk method showed negative result from all the isolates that cultured on International Streptomyces Project (ISP) 1 medium at 50oC and 40oC for 14 days. Screening by block agar method showed that two isolates, LC2-2 and LC2-6 gave positive result to Gram positive test bacteria. Morphologically, LC2-2 showed similarity to genus Bacillus, thus it’s not used for molecular identification. Molecular identification based on partial sequence of 16S rRNA gene with primer 785F and primer 802R showed that LC2-6 identified as Actinomadura keratinilytica (99%). Based on this research, it is suggested to do further study about the antimicrobial activity produced by LC2-6, because there is still no report about antimicrobial activity produced by Actinomadura keratinilytica.
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Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2014
S55886
UI - Skripsi Membership  Universitas Indonesia Library
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Adinda Rizki Putri
"Pleurotus ostreatus atau jamur tiram putih merupakan salah satu jamur tiram konsumsi dengan kandungan nutrisi dan senyawa bioaktif yang bermanfaat bagi kesehatan, salah satunya adalah kandungan enzim lakase yang berpotensi sebagai senyawa agen antikanker. Enzim lakase memiliki tingkat aktivitas yang tinggi dalam menghambat proliferasi sel kanker. Salah satu gen yang dapat mengkode enzim lakase pada P. ostreatus adalah gen POXA1b. Gen POXA1b dipilih karena memiliki tingkat ekspresi gen paling tinggi pada tiga fase hidup (miselia, primordia, dan tubuh buah). Saat ini belum pernah dilakukan desain primer, isolasi, dan optimasi primer dalam amplifikasi gen POXA1b dari P. ostreatus yang dibudidayakan di Indonesia. Oleh karena itu, penelitian ini bertujuan mendesain tiga pasang primer secara in silico, mengisolasi gen POXA1b dari tubuh buah P. ostreatus dan amplifikasi gen POXA1b dari P. ostreatus yang dibuktikan dengan analisis hasil sekuensing. Penelitian ini diawali dengan desain primer gen POXA1b yang dilakukan dengan bantuan laman NCBI, PrimerBLAST, dan perangkat lunak Snapgene, kemudian persiapan sampel dan isolasi DNA dari tubuh buah P. ostreatus. Hasil isolasi DNA diukur konsentrasi dan kemurniannya dengan spektrofotometer. Tahapan amplifikasi gen target dilakukan dengan teknik PCR. Hasil amplifikasi PCR divisualisasikan dengan elektroforesis gel agarosa lalu dilakukan analisis data. Konsentrasi isolat DNA dari tubuh buah P. ostreatus yang dihasilkan senilai 10,5—40,5 ng/nL dengan kemurnian senilai 1,694—2,189. Amplifikasi gen POXA1b dengan pasangan primer poxa1b-3F-A dan poxa1b-3R-A pada suhu annealing 59°C dan 60°C menghasilkan amplikon 244 bp. Berdasarkan hasil tersebut, desain primer, isolasi, dan optimasi primer pada amplifikasi gen POXA1b dari P. ostreatus hasil budidaya di Indonesia berhasil dilakukan dengan persentase homologi terhadap gen POXA1b dari P. ostreatus sebesar 100%.

Pleurotus ostreatus or white oyster mushroom is an edible oyster mushroom containing nutrients and bioactive compounds that are beneficial to health, one of them is the laccase enzyme content which has the potential as an anti-cancer agent. Laccase enzyme has a high level of activity in inhibiting the proliferation of cancer cells. One of the genes that encode the laccase enzyme in P. ostreatus is the POXA1b gene. The POXA1b gene was chosen because it has the highest gene expression in three life phases (mycelia, primordia, and fruit body) compared to other phenol oxidase genes. Currently, there has never been primer design, isolation, and primer optimization in the amplification of the POXA1b gene from P. ostreatus cultivated in Indonesia. Therefore, this study aims to design three pairs of primers in silico, isolate the POXA1b gene from fruiting bodies of P. ostreatus, and amplify the POXA1b gene from P. ostreatus as proven by analysis of the sequencing results. This study began with the design of POXA1b gene primers which was carried out with the NCBI website, PrimerBLAST, and Snapgene software, then sample preparation of P. ostreatus samples and DNA isolation from P. ostreatus fruiting body. The results of DNA isolation were measured for concentration and purity with a spectrophotometer. The stages of target gene amplification were carried out by PCR technique. The results of amplification were visualized by agarose gel electrophoresis and then data analysis was performed. The concentration of the DNA isolates from P. ostreatus fruiting bodies produced an average range of 10,5—40,5 ng/nL with a purity value of 1,694—2,189. The POXA1b gene amplification with primer pairs poxa1b-3F-A dan poxa1b-3R-A at annealing temperature of 59°C dan 60°C produced amplicons in the range of 200—300 base pairs. Based on the results, primer design, isolation, and the primer optimization of the amplification of the POXA1b gene from the P. ostreatus cultivated in Indonesia were successfully with a homology percentage to the POXA1b gene from P. ostreatus of 100%."
Depok: Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Indonesia, 2023
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UI - Skripsi Membership  Universitas Indonesia Library
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