本研究从含有伪狂犬病病毒(Pseudorabies virus，PRV)Ea株gD基因BamHI 6.6Kb片段的重组质粒pUCB7中分别亚克隆gD基因的上、下游片段，并对上游片段进一步改造，去掉gD基因上游的非编码序列，构建了含完整gD基因编码区的重组质粒pBRgDI，并利用基因内部的限制性内切酶位点，用内切酶KpnI和SalI酶切分析，证实了gD基因的可靠性和完整性。同时构建了测序质粒pSKgDSB和pSKgDS，并用双脱氧终止法进行测序。将测序结果与国外的Rice毒株进行比较，发现Ea株gD基因核苷酸序列有多处点突变和一处插入突变，在编码氨基酸残基水平上也有差异。%Glycoprotein gD gene of pseudorabies virus(PRV) Ea strain was cloned by means of restriction endonuclease digestion from plasmid pUCB7，which contained BamHI 6.6 kb fragment of the virus genome．For convenient for further study，the restriction endonuclease sites of EcoRI and BamHI were inserted in the up-and down-streams of gD gene encoding region，respectively．The reliable and integrated of gD gene was identified by cut with KpnI and SalI．The whole gene nucleotide sequence was also determined by dideoxygen termination and compared with those of PRV Rice strain．The results showed that there were multipile sitemutations and one insert-mutation in the encoding sequence of Ea’s gD gene．And the diversity of amino acid residues were also existing in glycoprotein gD of PRV Ea strain．
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