丝瓜籽核糖体失活蛋白基因Lufiin-a

目的探讨丝瓜籽核糖体失活蛋白基因Luffin—a原核表达载体的构建方法。方法从Pubmed中查到Luffin—a的mRNA全序列，用RT—PCR的方法从未成熟的丝瓜种子中克隆Luffin—a基因，T—A克隆后鉴定核苷酸序列，构建原核表达载体pET-15b（＋）-Luffin—a，转化大肠杆菌BL21（DE3），并经IPTG诱导表达；SDS-PAGE鉴定Luffin—a表达产物。结果克隆出Luffin—a基因，IPTG诱导后大肠杆菌培养液和超声破碎的菌体沉淀中都有Luffin—a表达产物。结论本研究成功构建了Luffin—a原核表达载体。

Construction of prokaryotic expression vector for ribosome inactivating protein gene Luffin-a of Luffa seeds

Objective To investigate the method of construction of prokaryotic expression vector for ribosome inactivating protein gene Luffin-a of Luffa seeds. Methods Whole mRNA sequence of Luffin-a was found from Pubmed database, Luffin-a gene from immature Luffa seeds was cloned by RT-PCR method, and the sequence of nucleotides was identified after T-A clone, then the prokaryotic expression vector of pET-15 b （＋）-Luffin-a was constructed, the vec tor was transfected into E. coli BL21 （DE3）, and the expression was inducted by IPTG. Luffin-a expression product was identified by SDS-page. Results Luffin-a gene was cloned. Luffin-a expression product was detected in the supernatant of E. coli culture solution and ultrasonic broken bacteria after IPTG induction. Conclusion This study successfully constructs prokaryotic expression vector of Luffin-a.