中国免疫学杂志2012，Vol.28Issue(3)：200-204,5.DOI:10.3969/j.issn.1000-484X.2012.03.003
小鼠B7-H4慢病毒表达载体的构建及对脾细胞增殖的影响
Construction of mouse B7-H4 lentivirus expression vector and its effect on the proliferation of mouse splenocytes
摘要
Abstract
Objective: To construct lentivirus expression vector of mouse B7-H4 and investigate its effect on the proliferation of mouse splenocytes. Methods :B7-H4 gene was cloned from normal lung tissue of BALB/c mouse. mB7-H4-EGFP vector was constructed. The mB7-H4-EGFP fusion gene was spliced out and ligated into lentivirus expression vector pCDHl-MCS1-EF1-puro. The resulting plasmid was designated pCDHl-mB7-H4-EGFP. The control vector pCDHl-Control-EGFP was constructed with a DNA oligonucleotide containing initiating Met and a multiple cloning site to replace mB7-H4. pCDHl-mB7-H4-EGFP or pCDHl-Control-EGFP was co-trans-fected into 293T cells with packaging plasmid mix by Iipofectamine? 2000 reagent. The supernatant of the cultured 293T cells was collected and used to infect 293T cells. MTT assay was used to assess the proliferation of BALB/c, C57BL/6 or BALB/c: C57BL/6(1: 1) mouse splenocyte that was co-cultured with mB7-H4 or control protein expressing 293T cells. Results: mB7-H4 was successfully cloned. Sequence analysis showed two nucleotides were different from those reported mB7-H4 sequence in GenBank, whereas their deduced amino acid sequences were consistent. mB7-H4 lentivirus expression vector were successfully constructed. mB7-H4-EGFP was expressed on the surface of infected 293T cells, and could inhibit the proliferation of BALB/c, C57BL/6 or BALB/c: C57BI/6(1: 1) mouse splenocyte. Conclusion;The mouse B7-H4 lentivirus expression vector was successfully constructed, and the functional mB7-H4 protein was detected on the surface of infected 293 T cells.关键词
B7-H4/慢病毒载体/293T细胞Key words
B7-H4/Lentiviral vector/ 293T cell分类
医药卫生引用本文复制引用
李丽,冯俊,王朝莉,敬保迁,胡为民..小鼠B7-H4慢病毒表达载体的构建及对脾细胞增殖的影响[J].中国免疫学杂志,2012,28(3):200-204,5.基金项目
本文为四川省科技厅应用基础课题(No.04JY029-014-2) （No.04JY029-014-2）
