Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Chinese Journal of Biotechnology》 2002-03
Add to Favorite Get Latest Update

Study on Construct and Expression of Synthetic Genes Encoding Spider Dragline Silk in Escherichia coli

LI Min+* ZHANG Wen-Xian HUANG Zhi-Hua HUANG Jian-Kun(College of Biological Engineering , Fujian Normal University, Fuzhou 350007, China)  
Dragline spider silk produced from Nephilia clavipes major ampullate is a natural fibrous protein with specific me- chanical properties such as high tensile strength and elasticity. Synthetic gene monomer encoding recombinant spider silk protein, based on the known repetitive protein sequence and partial cDNA sequence of dragline silk, was constructed and expressed. DNA monomer sequences were multimerized to encode high molecular weight synthetic spider silks using a “head-to-tail” construction strategy. Multimer was cloned into pET30a(+), a prokaryotic high potency expression vector, and induced with IPTG. The protein from 8-unit repeat was produced in Escherichia coli at levels up to 20 mg/L. The protein was easily purified with high recovery by using an metal ion affinity chromatography and purity was over 90%. The results of SDS-PAGE and Western blot suggested that the mass of the expression product was about 37kD. This value and amino acid analysis were consistent with those of theoretic calculation.
【Fund】: 福建省重大科技项目资助 (No.2 0 0 1F0 0 6 )~~
【CateGory Index】: Q78
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved