|Table of Contents|

Prokaryotic Expression System Construction of the Tufted Deer(Elaphodus cephalophus)AIF-1(PDF)

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

Issue:
2014年02期
Page:
85-
Research Field:
生命科学
Publishing date:

Info

Title:
Prokaryotic Expression System Construction of the Tufted Deer(Elaphodus cephalophus)AIF-1
Author(s):
Zhou Nannan; Yang Zhen; Luo Liyun; Song Fei; Bai Min; Cao Xiangrong
School of Life Sciences,Nanjing Normal University,Jiangsu Key Laboratory for Molecular and Medical Biotechnology,Nanjing 210023,China
Keywords:
Tufted deer(Elaphodus cephalophus); AIF-1; prokaryotic expression
PACS:
Q28
DOI:
-
Abstract:
The TdAIF鄄1 cDNA was cloned from the testis cDNA library of the Tufted deer(Elaphodus cephalophus) and analyzed by bioinformatic methods. Primers were designed according to cDNA sequence to clone the gene. The gene was cloned into pMD19鄄T vector for sequencing. The right sequence was digested by restriction enzyme and subcloned into the expression vector pET鄄28a( +). After transformed into E. coli BL21( DE3),the recombinant plasimid was induced to express by IPTG. The E. coli BL21(DE3)expressed recombinant protein was broken by ultrasonic to show whether the re鄄 combinant protein was soluble or not. Lastly,the soluble protein was purified. The recombinant protein was analyzed and identificated by SDS-PAGE electrophoresis and Western blot. Analysis of sequence showed that the TdAIF-1 cDNA contained a 438 bp op-n reading frame encoding 145 amino acids. The recombinant plasimid was correctly constructed according to sequencing and restriction enzyme analysis. The recombinant protein was about 20 kD and soluble mainly. When the recombinant protein was purified,using elution buffer containing 50 mmol/ L or 100 mmol/ L imidazole could get purified protein. The TdAIF-1 Prokaryotic Expression System was constructed successfully and recombinant protein was obtained,which was helpful for the future study of its biological function.

References:

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Last Update: 2014-06-30