摘要
目的：进一步了解人醛糖还原酶相似基因（aldose reductase like-1，ARL-1）的功能及其生物学活性，为制备特异性的ARL-1抗体作准备。方法：用基因重组技术构建PQE-ARL-1原核表达载体，在M15大肠杆菌中进行表达，然后利用Ni+-NTA琼脂柱纯化ARL-1蛋白。 结果：经酶切鉴定，PQE-30表达载体上已插入了ARL-1基因，PQE-ARL-1在大肠杆菌中表达产物人重组ARL-1 蛋白经纯化后进行SDS-PAGE电泳，成一条蛋白带，分子量为37.7×103，表达产物约占全菌蛋白的25%，定量测得ARL-1蛋白的浓度为100mg/L。 结论： PQE-ARL-1重组质粒的构建及ARL-1蛋白的制备，为深入研究ARL-1的功能及制备特异性的ARL－1抗体奠定了坚实的基础。
Objective: To further understand the function and biological activity of aldose reductase like-1 (ARL-1), and to provide the basis for the preparation of the specific antibody against ARL-1. Methods: The ARL-1 cDNA was cloned into procaryotic expression vector PQE-30 and the recombinant ARL-1 expressed in E.coli M15; then Ni+-NTA agarose columns were used to purify the ARL-1 protein. Results: With the digestion of the enzymes, we identified that the ARL-1 gene was inserted into the procaryotic expression vector PQE-30. The expression products of PQE-ARL-1 (human recombinant ARL-1) showed a single protein band on SDS-PAGE. The molecular weight of ARL-1 was approximately 37.7×103 and the expression level was about 25% of the total bacterial protein. The concentration of ARL-1 protein was about 100mg/L. Conclusion: The construction of the recombinant plasmid PQE-ARL-1 and the preparation of the protein ARL-1 have laid a solid foundation for further studying the function of ARL-1 and preparing the specific antibody against ARL-1.
出处
《中华肝脏病杂志》
CAS
CSCD
2000年第6期364-366,共3页
Chinese Journal of Hepatology
基金
国家自然科学基金!(39770863)
湖南省自然科学基金![湘科计字(97)37号-3]
