摘要：首次对犬瘟热病毒(CDV)大熊猫(GP)毒株附着或血凝蛋白(H)基因进行了序列测定并与疫苗株Onderstepoort进行了比较。我们设计合成了4对引物，对GP株进行了RT-PCR扩增与测序。H蛋白基因全长为1 946 bp，开放阅读框架(ORF)始于21-23位的ATG，终止于1 842-1 844位的TGA，编码607个氨基酸，该基因序列已被GenBank收录。将GP毒株与GenBank中疫苗弱毒株Onderstepoort进行比较，二者核苷酸序列的同源性为91.4%，推导的氨基酸序列的同源性为90.2%，GP和Onderstepoort株H蛋白的半胱氨酸残基数目均为12个且相对位置不变；疏水性有一定的变化，但推测的穿膜区位置(约35-55位氨基酸)是一致的；Onderstepoort株的H蛋白潜在的N-联糖基化位点为4个，GP株H蛋白为9个，糖基化位点的不同可能对GP株H蛋白的抗原性产生影响。

中图分类号：S852.65，Q78文献标识码：A

文章编码：1003-5125(2000)03-0291-06

Nucleotide sequecnce Analysis of the Haemagglutinin Protein

Gene of Giant Panda Strain of Canine Distemper Virus

LI Jin-zhong，XIA Xian-zhu，YU Chun，

FAN Quan-shui，HUANG Geng，QIU Wei，YEN Zhen

(The Veterinary Institute, University of Agriculture and Animal Science, Changchun, 130062, China)

HE Hong-bin

(Department of Biological Science, Northeastern Agricultural University, Harbin 150030, China)

Abstract: The sequence analysis of the haemagglutinin (H) protein gene of giant panda strain of canine distemper virus (CDV) from giant panda in China was conducted and compared with Onderstepoort vaccine strain. Four pairs of primers were designed and synthesized. The PCR products amplified from GP strain were directly sequenced after they were purified. There are 1 946 nt of the complete H gene. The H gene of the GP strain showed that the largest open reading frame (ORF) encoding a protein of 607 amino acid residues started at the ATG at position 21-23 and extended to a termination codon (TGA at position 1 842-1 844). The deduced amino acid sequence of the H protein of the GP strains was compared with the CDV vaccine Onderstepoort strain extracted from the GenBank database. The sequence of the H gene and deduced H protein of the GP strain showed an identity of 91.4% and 90.2% with Onderstepoort strain, respectively. All the 12-cysteine residues found in the GP strain H protein were conserved in Onderstepoort strain. Although predicted hydrophilicity of the GP strain differed from that of the Onderstepoort strain, the predicted transmembrane position at 35-55 amino acid residues was conserved in both strains. Out of the 4 potential N-linked glycosylation sites of Onderstepoort H protein, two were located in the N-terminal 2/3 of the protein (one located in the putative cytoplasmic part and is probably not used), while the remaining 2 sites were located in the C-terminal 1/3 of the protein. Four of the 9 potential N-linked glycosylation sites of GP H protein were conserved in the Onderstepoort, but the other five sites are unique to GP, the extra glycosylation sites may influence antigenicity of the GP.

Key word: Giant panda; Canine distemper virus; Haemagglutinin gene sequence analysis

基金项目：军队“八五”医药卫生基金重点课题(91-0205)

作者简介：何洪彬(1967年-)，男，汉族，黑龙江省讷河市，讲师，博士，主要从事分子病毒学研究。

参考文献

［1］张振兴，高素兰，徐福南，等.一起中国熊猫瘟热病的实验报告［J］.畜牧与兽医，1983，(4)：3-9

［2］ 胡桂学，夏咸柱，李金中，等.大熊猫犬瘟热致死性感染的首次报告［C］.见：第七届全国养犬学和第三届广东小动物医学联合会学术研讨会论文集，1997，172-174

［3］ Iwatsuki K, Miyashita N, Yoshida M, et al. Molecular and phylogenetic analyses of the haemagglutinin (H) proteins of field isolates of canine distemper virus from naturelly infected dog ［J］. J Gen Virol, 1997,78:373-380

［4］ Kovamess J, Blixenkrone-Moller M, Norrby E. The nucleotide and predicted amino acid sequence of the attachment protein of canine distemper virus ［J］. Virus Research, 1991,19:223-234

［5］ Chomczynski P and Sacchi N. Single stepo metheod of RNA isolation by acid guanidinium thiocyanate-phenol-choloroform extraction ［J］. Analytical Biochemistry, 1987,162:156-159

收稿日期：1999-05-31，修回日期：1999-10-18