Cyanovirin-N蓝藻的筛选及其分子鉴定

Screening and Molecular Identification of Cyanobacteria Including Cyanovirin-N Gene,a HIV-inactivating Protein

目的:从实验室培养的自生蓝藻中筛选出含有能编码抗病毒蛋白(cyanovirin-N,CV-N)基因的藻株并对其进行分子鉴定。方法:应用PCR技术和自行设计的特异性引物从80株蓝藻中筛选目标藻株,通过克隆、测序后,进行blast比对,经在线生物信息学软件预测该抗病毒蛋白CV-N的一级,二级,三级结构,并推测该基因序列编码的蛋白的抗病毒潜能。结果:筛选出藻株RZHA4,其所含的CV-N基因与已报道的CV-N基因有97%的相似性。结论:经16S rRNA-PCR和序列分析,该藻株与蓝藻Nostocaceae有99%的相似性,为一在实验室常规培养条件下生长良好的念珠藻(Nostoc),有可能成为新的CV-N生产藻株。

Objective：To screen a cyanobacterial isolate containing the gene encording Cyanovirin-N（CV-N） from free-living cyanobacterial strains cultured in the lab and to classify it molecularly.Method：Total 80 the algal strains/isolates were screened for CV-N gene using PCR with the primers designed specifically.The PCR product was cloned,sequenced,aligned through Blastn/Blastp and then predicted for the primary,secondary,and tertiary structural characteristics of the protein and potential functions of the putative Cyanovirin-N using the bioinformatic softwares on line.The target isolate was identified using 16S rRNA-PCR and sequence alignment.Result：A isolate RZHA4 carrying CV-N gene which had 97% similarity with the corresponding gene in GenBank.Conclusion：The RZHA4 was identified to belong to Nostocales due to 99% similarity on 16S rRNA sequence,being probably a new candidate for production of CV-N.Its potential function of antiviruses was speculated as well.