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In order to obtain three isoforms of apolipoprotein E (apoE), the eDNA encoding apoE3 was obtained by RT-PCR from normal human liver tissue, Site-directed mutagenesis was used to obtain the cDNAs encoding apoE2 and apoE4 isoforms. The 3 cDNAs were subcloned into vector pGEM-3Z and verified by DNA sequencing. The expression recombinant which can express the target protein as a (His) 6-tagged fusion was constructed by subcloning apoE eDNA into vector pTT-PL, The purified proteins were gained by Ni-NTA column, The SDS-PAGE results revealed the 6 His fusion proteins (apoE2, apoE3 and apoE4) were correctly expressed and purified successfully. 相似文献
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