脑钠肽重组蛋白的表达及生物学活性的研究

Brain Natriuretic Peptide Recombine Proteins Expression and Activity Researchment

目的比较脑钠肽（BNP）连接在不同载体中表达的BNP成熟肽蛋白和BNP前体蛋白的生物学活性差别。方法采用分子生物学技术构建重组质粒PGEX-20T—BNP32，PGEX-20T—BNP108,B11-BNP32和B11—BNP108分别对其进行PCR、双酶切和测序鉴定，然后将已测序鉴定的包含四种重组质拉的工程菌，转化至大肠埃希菌BL21菌中表达BNP成熟肽蛋白和BNP前体蛋白，并用ELISA法检测4种蛋白的生物学活性作用。结果在大肠埃希茵中表达的成熟肽蛋白BNP。2和前体蛋白BNP108经过纯化、复性后具有生物学活性。结论PGEx-20T—BNP52表达的成熟肽蛋白BNP32的生物学活性最强，为下一步单抗制备的优势蛋白。

Objective The difference between the biological activity of BNP mature polypeptides and its precursors was ex- amined in this paper. Methods Recombinant plasmid PGEX-20T-BNPa2,PGEX-20T-BNP108,B11-BNP32 and B11-BNP108 were constructed by molecuIar biological methods,which were identified by PCR ,restriction enzyme digestion and sequence analysis. Those four recombinant plasmids,identified by sequencing, were transformed into E. coli BL 21 and expressed BNP mature polypeptides and their corresponding precursors,whose biological activity was determined by EIA. Results BNP mature polypeptides BNP^2and its precursor BNP108 which were expressed in colon bacillus were reactive after purifi- cation and renaturation. Coaclusion BNP mature polypeptides which is expressed by PGEX-20T-BNP32 is most reactive and will be superior for inducing BNP-mAb in next step.