Seal B S
Virology Swine Research Unit, U.S. Department of Agriculture, Ames, IA 50010.
Virus Res. 1994 Jul;33(1):39-53. doi: 10.1016/0168-1702(94)90016-7.
Genomic variability within the capsid protein gene of feline calicivirus (FCV) was evaluated among different isolates using hybridization analysis and enzymatic viral nucleic acid amplification. Total infected cell RNA was first hybridized with cDNA clones generated to the capsid gene of the FCV isolates CFI/68, 255, LLK, NADC and KCD. Field isolates of FCV were categorized by hybridization with capsid gene cDNA from the reference strains. Isolates that did not hybridize were positive by Western blot using a cross-reactive cat polyclonal FCV CFI/68 capsid protein antiserum. Using previously published sequence information, oligonucleotide primers were generated based on conserved sequences surrounding the hypervariable capsid protein gene regions. Analysis of the FCV capsid protein gene hypervariable regions was completed by sequencing products of FCV nucleic acid amplified by reverse transcription and polymerase chain reaction. From these data, amino acid substitutions in the hypervariable regions of the capsid protein were identified for those isolates that did not hybridize with the original cDNA clones. An association between phylogenetic relationships and serum neutralization was established among FCV isolates examined.
利用杂交分析和酶促病毒核酸扩增技术,对不同猫杯状病毒(FCV)分离株的衣壳蛋白基因内的基因组变异性进行了评估。首先将总感染细胞RNA与针对FCV分离株CFI/68、255、LLK、NADC和KCD的衣壳基因生成的cDNA克隆进行杂交。通过与参考菌株的衣壳基因cDNA杂交,对FCV的野外分离株进行分类。未杂交的分离株通过使用交叉反应性猫多克隆FCV CFI/68衣壳蛋白抗血清的蛋白质免疫印迹法检测呈阳性。利用先前公布的序列信息,基于高变衣壳蛋白基因区域周围的保守序列生成寡核苷酸引物。通过对逆转录和聚合酶链反应扩增的FCV核酸产物进行测序,完成了FCV衣壳蛋白基因高变区域的分析。从这些数据中,确定了那些未与原始cDNA克隆杂交的分离株在衣壳蛋白高变区域的氨基酸取代情况。在所检测的FCV分离株之间建立了系统发育关系与血清中和之间的关联。