中国兽医科学2023，Vol.53Issue(12)：1531-1536,6.DOI:10.16656/j.issn.1673-4696.2023.0202
牛G3BP1蛋白的原核表达及其对牛cGAS生物酶催化活性的影响
Prokaryotic expression of bovine G3BP1 protein and its influence on enzymatic activity of the bovine cGAS
摘要
Abstract
To investigate the effect of bovine G3BP1(bG3BP1)on the enzymatic activity of DNA recog-nition receptor cGAS,the total RNA from bovine blood lymphocytes was extracted and reversely tran-scribed into cDNA by RT-PCR and the bG3BP1 gene were cloned.Subsequently,the prokaryotic expression vector pET-28a-SUMO-bG3BP1 was constructed and transformed into competent E.coli cells.The recombi-nant protein bG3BP1 was purified by nickel column affinity chromatography and purified further after removing SUMO label proteins to reach high purity.Finally,the bG3BP1 was used for the enzymatic reac-tion of bovine cGAS(bcGAS)in vitro,and the product of 2'3'-cGAMP was detected by ELISA.The results showed that soluble bG3BP1 protein promoted the enzymatic reaction of bcGAS in vitro and increased the yield of the second messenger molecule 2'3'-cGAMP,which provided technical methods and research ideas for the large-scale production and application of 2'3'-cGAMP.关键词
牛cGAS/牛G3BP1/原核表达/第二信使分子/酶促反应Key words
bovine cGAS/bovine G3BP1/prokaryotic expression/second messenger/enzymatic activity分类
农业科技引用本文复制引用
田慧慧,何小兵,陈国华,高真贞,韦雨松,俞宗佑,房永祥,米晓云,景志忠..牛G3BP1蛋白的原核表达及其对牛cGAS生物酶催化活性的影响[J].中国兽医科学,2023,53(12):1531-1536,6.基金项目
新疆动物疫病研究重点实验室开放基金项目(2023KLB003) （2023KLB003）
甘肃省自然科学基金项目(20JR10RA018,17JR5RA325) （20JR10RA018,17JR5RA325）
