Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (22): 4808-4815.doi: 10.3864/j.issn.0578-1752.2013.22.019

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Identification and Subcelluar Localization of Bmintegrin αPS3 from Silkworm (Bombyx mori)

 TAN  Juan, ZHANG  Kui, XU  Man, CHEN  Si-Yuan, CUI  Hong-Juan   

  1. State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing 400716
  • Received:2013-04-23 Online:2013-11-15 Published:2013-07-10

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Abstract: 【Objective】Bmintegrin αPS3 was cloned, analyzed, and overexpressed either in Escherichia coli or silkworm cell line to provide a theoretical basis for further studying the function of Bmintegrin αPS3 in silkworm.【Method】The Bmintegrin αPS3 cDNA including the complete ORF sequence was cloned by RACE method. The spatial-temporal expression profile of Bmintegrin αPS3 was investigated by RT-PCR. Bmintegrin αPS3 protein was overexpressed using prokaryotic expression. In addition, Bmintegrin αPS3 was constructed into a transit expression vector, and transfected into silkworm embryonic cell line to analyze the subcellular location of Bmintegrin αPS3.【Result】Bmintegrin αPS3 cDNA consisted of 2 895 bp, and encoded 965 amino acids containing the conserved integrin alpha domains, which was predicted to be a transmembrane protein. Bmintegrin αPS3 protein from E. coli was overexpressed and purified using affinity chromatography, and the purity reached more than 99%. In addition, the overexpressed Bmintegrin αPS3 protein in embryonic cell line was located in cytoplasm and membrane. RT-PCR results showed that Bmintegrin αPS3 mRNA was specifically and continually expressed in silkworm hemocytes.【Conclusion】The Bmintegrin αPS3 cDNA was cloned and its expression patterns in silkworm were analyzed. The subcelluar localization of recombinant Bmintegrin αPS3 protein was investigated in silkworm embryonic cell line. The Bmintegrin αPS3 protein was overexpressed in E. coli and purified for further antibody preparation.

Key words: Bmintegrin αPS3 , cloning , spatial-temporal expression , prokaryotic expression , subcellular localization

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