Study on Cloning, Expressing and Bioactivity of S-8300′s cDNA Fragment
HUANG Feng-jie, L Li-li, QIAN Jing, WU Wu-tong*
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School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
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History+
Received
Revised
Published
2012-01-01
2012-01-01
2012-01-01
Issue Date
2010-01-15
Abstract
OBJECTIVE Study on cloning, expressing and bioactivity of a new peptide S-8300. METHODS A deduced oligonucleotide primer was designed according to the N-terminal amino acid residues of the new peptide S-8300. One cDNA segment was amplified from the total RNA of regenerated hepatic tissue using the technique of RT-PCR, and its nucleotides were 342 bp. RESULTS According to the result of sequence analysis and derived amino acids sequence, this fragment′s amino acid residues had similarity with S-8300 at the end of N-terminal. In this study, the cDNA encoding S-8300 was inserted into the downstream of signal peptide PelB to construct the secretory plasmid pET22b-S which was transformed into Escherichia coli BL21(DE3) competent cells. Then the prokaryotic secretory expression strain pET22b-S /BL21(DE3) was constructed. When the induced with 0.4 mmol·L-1 IPTG at 25 ℃, the recombinant protein rS-8300 was secreted into the periplasm of the engineering strain in a soluble form effectively. CONCLUSION It had the same activities with natural products which could recover the damage of NIT-1 cell after exposed to the toxin of streptozocin.
HUNG Feng-jie;L Li-li;QIN Jing;WU Wu-tong.
Study on Cloning, Expressing and Bioactivity of S-8300′s cDNA Fragment[J]. Chinese Pharmaceutical Journal, 2010, 45(2): 93-97
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