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Construction of Follicle-specific Expression Vector of Spider Dragline Silk Protein and Screening of the Transgenic Cell Lines

WANG Hai-tao,MENG Fan-hua,FANG Jun,ZHOU Huan-min   

  1.  (Key Laboratory of Biological Manufacturing, College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018China)

  • Received:2013-11-18 Online:2014-06-20 Published:2014-07-24

Abstract: This study aimed at getting transgenic cell lines which could express spider dragline silk protein in the hair follicles specifically. By synthesizing spider dragline silk protein gene monomer artificially according to the published cDNA fragments of Nephila clavata spider, got the diploid (2S) by doubling, then constructed hair follicles specific expression vector pK-2S using pEGFP-N1 as original vector and transfected Xinjiang fine wool merino sheep fetal skin fibroblasts. Make use of PCR to detect positive clones at DNA and RNA level.The sequencing result showed correct sequence after gene synthesis,after enzyme digestion of the vector, we got the correct band while we could amplify target gene from the genom and cDNA of positive clones. This study established cell lines of transgenic spider dragline silk like protein gene successfully while the gene had expressed at RNA level. Our study laid the foundation of making the spider dragline silk gene specifically expressed in the hair follicles of the sheep by transgenic technology.

Key words:

fine wool sheep; spider dragline silk protein gene; specially expressed in hair follicle