稀杯盔形珊瑚钙网蛋白基因的克隆及功能分析

钙网蛋白（calreticulin，CRT）是高度保守的内质网钙结合蛋白和分子伴侣，主要参与调节钙离子水平和指导蛋白质正确折叠。在这里，我们克隆稀杯盔形珊瑚（Galaxea astreata）的CRT基因，命名为Gacrt，并分析其促进细菌凝集的能力。Gacrt全长1792bp，其中5’端非翻译区(5''UTR)为77 bp，3’端非翻译区(3''UTR)为380bp，开放阅读框(ORF)1335bp，编码444个氨基酸残基，包含信号肽结构域、内质网检索信号序列（KDEL）、两条保守的钙网蛋白家族结构和一组三重复序列。通过构建原核表达系统得到CRT重组蛋白，纯化后的重组蛋白可以促进革兰氏阳性菌藤黄微球菌和革兰氏阴性菌大肠杆菌凝集，具有细菌凝集活性，支持GaCRT作为免疫相关分子参与机体对细菌防御的观点。

Cloning and functional analysis of the calreticulin gene from the scleractinian coral Galaxea astreata

Calreticulin(CRT) is a highly conserved Ca2+-binding protein and chaperone in the endoplasmic reticulum,which mainly participates in adjusting calcium level and directing proper conformation of proteins. Here, we cloned the crt gene of the scleractinian coral Galaxea astreata, named Gacrt, and analysed its ability to drive bacterial agglutination. The full-length Gacrt cDNA consisted of 1 792 nucleotides and contained a 77 bp 5'untranslated region(UTR), a 380 bp 3' UTR and a 1 335 bp open reading frame(ORF) that encoded a 444 amino acid protein. The deduced peptide possessed a signal peptide domain, an endoplasmic reticulum retrieval signal sequence(KDEL), two potential calreticulin family signature motifs and a set of triplicate repeats. We also found that the recombinant GaCRT protein could promote agglutination of both the Gram-positive bacterium Micrococcus luteus and the Gram-negative bacterium Escherichia coli. These results show that the GaCRT protein can enhance bacterial agglutination, hinting that GaCRT is an immune-relevant molecule involved in host defense against bacterium.