猪BPI蛋白重组载体的构建及真核表达

试验旨在构建猪BPI真核表达载体,获得猪源BPI重组蛋白.根据GenScript's CloneEZ^® PCR Cloning Kit试剂盒,将BPI编码序列克隆至pUC57载体上,酶切与测序鉴定无误后,将重组质粒pUC57-BPI转染293-6E细胞,并利用SDS-PAGE和Western blotting方法检测其表达水平.结果显示,本试验成功构建了猪源BPI蛋白真核表达载体pUC57-BPI,并在293-6E细胞培养上清中检测到猪源BPI重组蛋白的表达.该猪源BPI重组蛋白体外表达系统的建立为今后深入研究猪BPI的生物学功能以及猪抗菌蛋白的制备提供了试验基础.

Construction and Eukaryotic Expression of Porcine BPI Protein Recombinant Vector

In order to construction porcine bactericidal/permeability-increasing protein (BPI) eukaryotic expression vector and obtain porcine BPI protein recombinant vector. In this study, BPI coding sequence was cloned into vector pUC57 according to GenScript''s CloneEZ^® PCR Cloning Kit. After correct identification with digestion and sequencing, the recombinant plasmid pUC57-BPI was transfected to 293-6E cells, then the expression of the recombinant protein was detected by SDS-PAGE and Western blotting. The results showed that we successfully constructed the eukaryotic expression vector pUC57-BPI. Furthermore, porcine BPI recombinant protein was expressed in cultural supernatant of 293-6E cells in eukaryotic expression system, and the establishment of porcine BPI recombinant protein expression system in vitro provided the basis for the deep study of porcine BPI biological function and the preparation of porcine antibacterial protein.