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Abstrak :
Jamur tiram putih (Pleurotus ostreatus (Jacq.) Kumm 1871) diketahui mengandung senyawa bioaktif yang bermanfaat bagi kesehatan. Salah satunya adalah senyawa beta glukan (β-glukan) yang memiliki kemampuan imunomodulator dengan meningkatkan jumlah sel natural killer dan mendukung perkembangan respons imun. Senyawa β-glukan dikode oleh gen FKS, yaitu gen spesifik pada kompleks β-1,3-glukan sintase. Gen FKS terekspresi di fase miselia, primordia, bakal tubuh buah, dan tubuh buah dewasa P. ostreatus dan paling tinggi pada fase tubuh buah dewasa. Studi mengenai desain primer, isolasi, dan optimasi primer untuk amplifikasi gen FKS dari P. ostreatus yang dibudidayakan di Indonesia belum pernah dilakukan. Oleh sebab itu, penelitian ini dilakukan untuk mendesain dua pasang primer (FKS-A dan FKS-B) secara in silico, mengisolasi gen FKS dari tubuh buah P. ostreatus, dan optimasi primer untuk amplifikasi gen FKS. Penelitian diawali dengan desain primer yang dibantu oleh NCBI PrimerBlast dan Primer3Plus, kemudian DNA diisolasi dari tubuh buah P. osteatus. Konsentrasi dan kemurnian isolat DNA diukur menggunakan spektrofotometer. Gen target diamplifikasi dengan teknik PCR yang kemudian divisualisasikan dengan elektroforesis gel agarosa. Isolat DNA dari tubuh buah P. ostreatus memiliki konsentrasi senilai 2,803—22,616 ng/μL dengan rerata 14,819 ng/μL dan kemurnian di rentang 1,815—2,083. Pasangan primer FKS-A dan primer FKS-B berhasil mengamplifikasi dan menghasilkan amplikon berukuran 100—200 bp dan 300—400 bp yang diduga sebagai gen FKS. Hasil tersebut menunjukkan bahwa desain primer, isolasi dan optimasi primer untuk amplifikasi yang diduga sebagai gen FKS dari P. ostreatus berhasil dilakukan. ......White oyster mushroom (Pleurotus ostreatus (Jacq.) Kumm) is known to contain bioactive compounds that are beneficial to health. One of them is the beta glucan compound (β-glucan) which has immunomodulatory abilities by increasing the number of natural killer cells and supporting the development of the immune response. β-glucan compounds are encoded by the FKS gene, which is a specific gene in the β-1,3-glucan synthase complex. The FKS gene is expressed in the mycelia, primordia, young fruiting body, and mature fruiting body phases of P. ostreatus and is highest in the mature fruiting body phase. Studies on primer design, isolation, and primer optimization for FKS gene amplification of P. ostreatus cultivated in Indonesia have never been conducted. Therefore, this study was conducted to design two pairs of primers (FKS-A and FKS-B) in silico, isolate the FKS gene from the fruiting body of P. ostreatus, and optimize the primers for the amplification of the FKS gene. The research began with a primer design assisted by NCBI PrimerBlast and Primer3Plus, then DNA was isolated from the fruit body of P. osteatus. The concentration and purity of DNA isolates were measured using a spectrophotometer. The target gene was amplified by PCR technique which was then visualized by agarose gel electrophoresis. DNA isolates from the fruit body of P. ostreatus had concentrations of 2,803—22,616 ng/μL with an average of 14,819 ng/μL and purity in the range of 1,815—2,083 with an average of 1,978. Primer pairs of FKS-A and FKS-B successfully amplified and produced amplicons measuring 100-200 bp and 300-400 bp which are suspected to be FKS genes. The results showed that the primer design, isolation and primer optimization for the amplification of the suspected FKS gene from P. ostreatus were successfully carried out.

Abstrak :
ABSTRACT
Actin is a major component of the plant cytoskeleton, so all cells contain this protein. Actin is expressed constitutively and is involved in basic housekeeping functions required for cell maintenance. Because of this, it has been frequently used as an internal control to normalize changes in gene expressions analysis. Actually, the information of nucleotide sequence of actin gene of Jatropha curcas L. population IP-2P from Indonesia is not available yet. The objective of this research was to isolate, clone and characterize cDNA of actin genes of J. curcas IP-2P. Three partial actin gene sequences had been successfully isolated by PCR using total cDNA as template, and actin primer designed from conserved region of Arabidopsis thaliana. Nucleotide sequence analysis showed that the length of JcACT fragment is 610, 534, and 701 bp encoding 203, 177, and 234 amino acids respectively. Local alignment analysis based on mRNA sequences shows that JcACT fragment shares 98% similarity with actin mRNA of Hevea brasiliensis and 99% with actin mRNA of Ricinus communis. Based on deduced amino acid sequence, JcACT is 100% identical to acting from Prunus salicina, Gossypium hirsutum, and Betula luminifera. Even though these clones of cDNA are not completed yet, they can be used as reference in J. curcas L. gene expression analysis.

Abstrak :
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.