Construction of prokaryotic expression plasmids

CH Chen-Chen Huang
FL Fang-Rui Liu
QF Qiang Feng
XP Xin-Yan Pan
SS Shu-Ling Song
JY Ju-Lun Yang
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The anti-p21Ras scFv was constructed previously in our laboratory [30]. The RGD4C peptide (ACDCRGDCFCG) was developed with phage display technology [42]. The anti-p21Ras scFv gene and RGD4C gene were linked genetically and then inserted into the prokaryotic expression plasmid pET-28a (+) between the BamH I and Hind III sites. The pET-28a (+) expression vector contains two 6 × His tags to allow immobilized metal ion affinity purification. Recombinant plasmids were sequenced for identification (Qingke, China). Four prokaryotic expression plasmids were constructed: p-scFv, p-RGD4C-scFv, p-RGD4C-linker-scFv, and p-RGD4C-EGFP.

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