论文部分内容阅读
为探明S–位点受体激酶(SRK)上与类硫氧还蛋白1(THL1)作用的氨基酸区域以及两者间作用方式,依据SRKE1上功能域分布构建了两种SRKE1短截体原核表达质粒pGEX-SRKE1A和pGEX-SRKE1B,分别在大肠杆菌BL21中获得了可溶性表达。通过体外孵育检测蛋白质相互作用的方法对SRKE1A、SRKE1B与THL1相互作用进行了检测,结果表明SRKE1A和SRKE1B均可与THL1结合,明确了THL1与SRK相互作用并不依赖于SRK激酶活性。两类SRK等位序列间比对结果表明Cys在两类SRK间的保守性存在明显差异,推测两类SRK材料亲和性的差异可能与Cys保守性不同有关。
In order to elucidate the amino acid regions of S-site receptor kinase (SRK) interacting with Thioredoxin 1 (THL1) and the mode of action, two SRKE1 prokaryotic nuclei were constructed based on the functional domain distribution of SRKE1 The expression plasmids pGEX-SRKE1A and pGEX-SRKE1B were obtained in soluble form in E. coli BL21 respectively. The interaction between SRKE1A, SRKE1B and THL1 was detected by in vitro incubation. The results showed that both SRKE1A and SRKE1B could bind to THL1, which indicated that the interaction between THL1 and SRK did not depend on SRK kinase activity. The alignment of the two SRK alleles showed that there was a significant difference in the conservation of Cys between the two types of SRKs. It is speculated that the differences in the affinity between the two SRKs may be related to the differences in Cys conservation.