Cloning and Expression of Spider Dragline Silk Protein Gene in Escherichia coli and Eukaryotic Cells  

Danmei Liu1,2 , Fen Wang1 , Wenli Li1
1. School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, 116023, P.R. China
2. School of Agriculture, Eastern Liaoning University, Dandong, 118001, P.R. China
Author    Correspondence author
Molecular Entomology, 2011, Vol. 2, No. 1   doi: 10.5376/me.2011.02.0001
Received: 19 Nov., 2011    Accepted: 26 Dec., 2011    Published: 28 Dec., 2011
© 2011 BioPublisher Publishing Platform
This article was first published in Genomics and Applied Biology in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:

Liu et al., 2011, Cloning and Expression of Spider Dragline Silk Protein Gene in Escherichia coli and Eukaryotic Cell, Genomics and Applied Biology, 30(1): 16-20 (doi: 10.3969/gab.030.000016)

Abstract

With the extreme tensile strength and toughness, spider silk has a high application value in a wide range of medical and industrial. However, the application of spider silk was limited because of its non-domestication. So, we tried to obtain the spider silk protein by genetic engineering. In this study, the dragline silk gene (ASP), a 837 bp fragment was cloned from the genome of spider (Araneus ventricosus) by Nested PCR. And then it was ligated into pGEX-6p-1 prokaryotic expression vector and pGFP-N2 eukaryotic expression vector, named as pASG and pASN, respectively. The fusion protein GST-ASP was successfully expressed in pASG and characterized by Western blotting with anti-GST antibodies after induced at 16℃ for 24 h in E. coli. The fusion proteins ASP-GFP was also successfully expressed in pASN by the detection of green fluorescence after transfected in insect sf9 cells for 8 h, which indicated that the ASP gene was correctly expressed in E. coli and eukaryotic cells, respectively. This study provides a beneficial attempt to develop the pathway for yielding spider silk protein by genetic engineering.

Keywords
Dragline silk gene; Gene cloning; Prokaryotic expression; Eukaryotic expression
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