MOLECULAR CLONING AND NUCLEOTIDES SEQUENCE ANALYSIS OF G1 GENOME SEGMENT OF HANTAAN VIRUS Z10 STRAIN

摘要

The molecular cloning of the G1 genome segment of the Z10 strain of hantaan virus and analysis of the first gene data on the currently widely used Hantavirus vaccine strain in China are presented. The G1 genome segment of Z10 virus was amplified by the method of RT-PCR, and the products were cloned into the pGEM-T vector after identification and purification. The clone was sequenced by Sanger's dideoxy chain termination method. The 1449 bases of the Z10 G1 genome segment ,and the coding for 483 amino acids were determined. The base compositions for the G1 segment RNA determined from cDNA sequence information , were 20.6% A, 21.6% G, 18.8% C and 30.0% U. These values are similar to those of the 76/118, Lee and SR virus. As compared to the Z10 G1 segment, the sequence homology at the nucleotide level is 87%(76/118, type I), 86% (Lee, type I),86% (Hojo, type I), 67%(R22,type II), and 59%(K22, type III).The Z10 virus G1 protein amino acid sequence identity with other hantaan viruses (94-95% homology) was higher than that with Seoul type viruses (77-80%). More amino acid sequence heterogeneity between the Z10 and 76/118 was observed in the N terminal, especially the diverging cluster of ten amino acids at position 84 to 93 of the Z10 virus G1 protein. Conclusion:(1) The Z10 strain is one of the hantaan viruses. (2) The important region of the Z10 G1 segment was conservative. (3) Although substantial divergence of nucleotide sequences of the G1 genome segment was found between the Z10 strain and other type I Hantaviruses, relatively high amino acid sequence homology was shown among them. Thus, good immune protection could be obtained with the inactivated Meriones unguiculatus kidney cell vaccine against other strains of Hantaviruses.