伪狂犬病毒糖蛋白H基因片段的克隆、序列测定和分析

对伪狂犬病毒湖北地方株(PRVHB株)糖蛋白H基因片段进行了扩增、克隆、序列测定和分析,比较了该序列与PRVKa株及NIA-3株三者之间的同源性.结果显示,克隆片段长1396bp,G+c含量75.6%,包括糖蛋白H(gH)N端283个氨基酸编码区及上游调控序列和胸苷激酶(TK)C端136个氨基酸编码区.gH基因ORF上游调控区存在有TATA框和CAT框的真核启动子特征结构.PRVHB株gH基因片段序列与PRVKa株和N1A-3株相应序列的核苷酸同源性相同,为99.6%.推导的gH多肽N端第1～30位氨基酸组成的肽段富含疏水性氨基酸,具有真核信号肽的序列特征.

Cloning Sequencing and Structrual Analysis of the Glycoprotein H Gene of A Wild Chinese Pseudorabies Virus Strain

With pured PRV genome DNA as template, the part region of glycoprotein H gene and the upstream tk gene located with genomic BamHI fragments 11 and 16 was amplified by PCR, and its nucleotide sequence was determined by the dideoxy\|mediated chain termination method. A analysis of the nucleotide sequence and its deduced amino acid seqnence was performed with computer programs. The results revealed that the region of DNA sequenced was 1396bp and had the content of (G C) 75.6%. The nucleotide sequence comparison of gH gene showed that the homology of the nucleotide sequence of PRV HB strain with PRV Ka strain and NIA\|3 strain was both 99.6%. The deduced amino acid sequence of gH polypeptide exhibited a strongly hydrophobic region present at the N\|terminus at position 1~30 amino acids, and exhibited features typical for eucaroytic signal sequences.