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- W2351474377 abstract "Objective To express and detect the antigenicity of human laminin alpha4 LG3-4 module (hLNα4LG3-4) protein by gene engineering techniques. Methods The cDNA encoding hLNα4LG3-4 was amplified by RT-PCR from human placenta, then inserted into pMD-18T vector by T/A cloning and sequenced. Prokaryotic expression vector pET-28a-LG3-4 was constructed by recombinant DNA technique. The hLNα4LG3-4 fusion protein expressed in BL21(DE3)/pET system was identified by SDS-PAGE, purified by Ni-NTA resin, and assayed by Western blotting. Results The cDNA fragment of hLNα4LG3-4 was cloned successfully. While BL21 (DE3)/pET28a-LG3-4 bacterium was induced with IPTG, a new protein band with a relative molecular weight of 44 000 was shown on SDS-PAGE profile. hLNα4LG3-4 fusion protein of high purity (95%) was obtained and specific protein band was detected by Western blotting. Conclusion The hLNα4LG3-4 fusion protein was successfully expressed." @default.
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- W2351474377 date "2005-01-01" @default.
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- W2351474377 title "Cloning and expression of human laminin alpha4 LG3-4 module" @default.
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