Open AccessISOLASI KARAKTERISASI DAN UJI EKSPRESI GEN ALP1 DI ESCHERICHIA COLI DH5A
Author(s) -
Ni Nyoman Tri Puspaningsih,
Akhmaloka Akhmaloka,
Sofyan Hadi,
Yosephine Sri Wulan Manuhara,
Bambang Irawan
Publication year - 1999
Publication title -
berkala penelitian hayati (journal of biological researchers)/berkala penelitian hayati
Language(s) - English
Resource type - Journals
eISSN - 2337-389X
pISSN - 0852-6834
DOI - 10.23869/bphjbr.4.2.19992
Subject(s) - ecorv , ecori , restriction enzyme , insert (composites) , microbiology and biotechnology , recombinant dna , bamhi , escherichia coli , gene , chemistry , cloning (programming) , biology , biochemistry , materials science , computer science , programming language , composite material
Amylase gene (ALP1) insertion in YCp 50 cloning vector was detected by Direct Screening PCR. Result of Direct Screening PCR showed one transformants which assumed have amylase gene insert. Characterization of the recombinant DNA by Stu1 and EcoR1 restriction enzymes indicated nucleotide sequence of insert gene. Digestion by BamHI, Cla1, and EcoRV restriction enzymes showed only one band that assumed size of insert gene is about 1500 bp. Gene expression showed that amylase enzyme activity by using Somogyi-Nelson method was 88.0265 Unit. This activity was 10 percent higher than transformant control (Escherichia coli DH5a which content YCp50).