CXCL1基因重组慢病毒表达载体的构建及鉴定

目的：采用第2代慢病毒表达载体系统，构建CXCL1基因的慢病毒表达载体，摸索出较传统转染方法更为高效、可靠的转基因方法，为今后进一步的基因靶向治疗研究打下良好的基础。方法：首先扩增CXCL1全长序列，将其与慢病毒载体pWPI连接后测序，通过BLAST检索，鉴定慢病毒表达载体构建成功。将重组慢病毒质粒转染293T细胞，48h后荧光显微镜鉴定，慢病毒转染并包装成功。结果：重组慢病毒质粒CXCL1-pWPI测序结果经BLAST对比分析，与CXCL1基因的同源性达100％。结论：成功的构建了CXCLl基因的重组慢病毒表达系统。

CONSTRUCTION AND IDENTIFICATION OF CXCL1 RECOMBINANT LENTIVIRAL EX- PRESSION VECTOR

Objective： The second-generation lentiviral vector system was used to constructed CXCL1 gene lentiviral vector, and to explore the more efficient and reliable transgenic approach compared with tradi- tional transfection methods for the further study of gene therapy targeted. Methods： First full-length se- quence of CXCL1 was amplified by PCR, and was connected with the lentiviral vector pWPI, as well as was sequenced by BLAST search to identify that lentiviral vector was constructed successfully. The recombi- nant lentiviral vector was transfected to 293T cells and was identificated by fluorescent microscopy after 48 hours in order to comfirm lentiviral transfection and packaging successfully. Results： The sequencing re- suits of the recombinant lentiviral plasmids CXCLI-pWPI compared by BLAST analysis showed the CXCL1 gene homology of 100~/00. Conclusion： The CXCL1 gene recombinant lentiviral expression system was con- structed successfully, and that would be useful for CXCL1 gene function research.