The KLF3 gene was a potential genetic marker candidate due to its involvement in many biological processes such as cell proliferation, cell differentiation, and apoptosis. This study was conducted to identify the KLF3 gene polymorphism in the Indonesian FH cattle. A total of 302 individual cattle blood samples from West Java (n=138), Central Java (n=34), and East Java (n=130) provincies were used to obtainDNA samples. The DNA sequencing was performed using 9 samples (each location of sampling represented by 3 samples) for initial identification of the KLF3 gene polymorphismand followed by PCR-RFLP analysis using all DNA samples. There were two SNPs identified in the intron region of the KLF3 gene i.e. SNP g.59607486delC and SNP g.59607554A>G. The SNP g.59607554A>G could be detected using PCR-RFLP method with HpyCH4IV restriction enzyme and resulted three genotypes of the KLF3 gene (AA, AG, and GG) but its heterozygosity value was low in the Indonesian FH cattle. The AA genotype has the highest frequency (0.73), while the AGand GG genotypes frequency were 0.24 and 0.03, respectively. Information about the KLF3 gene polymorphism in this study can be considered for further study to analyse its association with milk and protein yield traits.
Gen proinsulin telah berhasil diinsersi pada galur Pichia pastoris X33, GS115 dan KM71H pada penelitian sebelumnya, namun masih memerlukan optimasi. Penelitian ini dilakukan untuk mengetahui ekspresi galur P. pastoris dan waktu induksi yang optimal dalam mengekspresikan proinsulin. Ekspresi proinsulin dilakukan pada suhu 20 °C selama 120 jam untuk pemilihan galur terbaik. Variasi lama induksi yang digunakan adalah 0, 24, 48, 72, 96, 120 dan 144 jam untuk mengetahui waktu induksi optimal. Kultur kemudian dilisis, dielektroforesis menggunakan Tricine SDS PAGE dan divisualisasikan dengan pewarnaan perak. Berdasarkan ketebalan pita yang terbentuk pada gel elektroforesis, galur X33-X2 menghasilkan proinsulin terbanyak dibandingkan GS115-G11 dan KM71H-K4.. Ekspresi proinsulin optimal pada 120 jam dan mengalami penurunan proinsulin pada waktu induksi 144 jam akibat akumulasi metanol. Galur X33-X2 merupakan inang terbaik dan waktu induksi 120 jam merupakan waktu induksi optimal dalam mengekspresikan proinsulin secara intraseluler. Hasil ini diharapkan dapat menjadi referensi dalam mengembangkan produksi insulin di Indonesia.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.