CAJ | 학술논문

目的：构建重组表达载体pGEX-脑源性神经营养因子（BDNF），探讨其原核表达BDNF的可行性。方法：大鼠BDNF进行密码子优化，人工合成BDNF的全基因序列，经回收、纯化、酶切后连接到pGEX原核表达载体中，将重组质粒pGEX-BDNF转化大肠杆菌BL21细胞中，筛选阳性克隆，经IPTG诱导表达后，利用ELISA方法鉴定其表达的活性。结果：PCR鉴定及测序显示BDNF序列完全正确，BDNF基因的克隆和表达载体构建成功，IPTG诱导表达目的蛋白经ELISA检测，其表达的目的蛋白具有一定的活性。结论：经密码子优化后经人工合成得到BDNF基因片段并成功构建pGEX-BDNF重组表达载体，具有一定的生物活性。
목적：구건중조표체재체pGEX-뇌원성신경영양인자（BDNF），탐토기원핵표체BDNF적가행성。방법：대서BDNF진행밀마자우화，인공합성BDNF적전기인서렬，경회수、순화、매절후련접도pGEX원핵표체재체중，장중조질립pGEX-BDNF전화대장간균BL21세포중，사선양성극륭，경IPTG유도표체후，이용ELISA방법감정기표체적활성。결과：PCR감정급측서현시BDNF서렬완전정학，BDNF기인적극륭화표체재체구건성공，IPTG유도표체목적단백경ELISA검측，기표체적목적단백구유일정적활성。결론：경밀마자우화후경인공합성득도BDNF기인편단병성공구건pGEX-BDNF중조표체재체，구유일정적생물활성。
Objective: To construct and identify the prokaryotic recombinant expression vector pGEX-BDNF. Methods:The BDNF sequence from NCBI was optimized. The BDNF sequence was synthesized and ligated into pGEX prokaryotic expression vectors after recovery, purification and enzyme digestion. The purified recombinant plasmid was transformed into the E. coli BL21 cells. The positive clones were selected by PCR .The protein activity was identified by ELISA after IPTG induction. Results:PCR results and sequencing analysis showed that BDNF gene was correctly cloned into the prokaryotic recombinant expression vector pGEX. The ELISA results showed that the expression protein has a certain activity after IPTG induction. Conclusion: The recombinant prokaryotic expression vector pGEX-BDNF was successfully constructed.