小鼠生精相关基因Vad1.2表达载体的构建及鉴定

摘要

Objective: To construct and characterize the Vad1.2 expression vector related to spermatogenesis in mice. Methods:Vad1.2 transcript was amplified from mouse testis by RT- PCR. The amplified product was inserted into pET15b vector. Then the Vad1.2 over- expression vector was transformed into E. coli BL21 (DE3) RIL competent cells and identified by endonuclease digestion and DNA sequencing. Results: Putative positive clones were verified by double restriction digestion, displaying the pET- 15b vector backbone and the insert with correct size. Conclusion:The Vad1.2 over expression vector has been successfully constructed, which is of significance for the functional study of Vad1.2 gene in spermatogenesis.%目的：构建小鼠生精相关基因Vad1.2表达载体并进行鉴定。方法：将小鼠睾丸组织中Vad1.2转录本进行RT- PCR扩增，通过DNA重组技术将其插入克隆载体pET15b中，将表达载体转化入大肠杆菌BL21（DE3）RIL感受态细胞，提取重组质粒并通过酶切及DNA测序进行鉴定。结果：随机挑取的克隆经双酶切鉴定，证实Vad1.2表达载体构建成功，插入基因序列正确。结论：成功构建了小鼠Vad1.2基因表达载体，为Vad1.2基因的进一步研究奠定了实验基础。