摘要
本实验为了评价表达新城疫病毒F基因重组鸡痘病毒(rFPV282E4-SF,rFPVLp-SF)的遗传稳定性,将其在鸡胚成纤维细胞连续培养20代。通过蓝斑来鉴定重组鸡痘病毒纯度,结果所有蚀斑100%变蓝。通过对第0、10和20代F基因扩增并进行序列测定,所有碱基和氨基酸序列均和原始转移载体序列完全一致,没有发生任何改变。随后又通过间接免疫荧光证实rFPV282E4-FS和rFPVLP-FS各个代次F基因的特异表达。所有结果均显示了上述重组鸡痘病毒具有良好的遗传稳定性,满足兽医生物制品的遗传稳定性要求。
In order to evaluate the genetic stability of two recombinant fowlpox viruses （rFPV282E4-FS, rFPV Lp-FS） expressing fusion protein gene of ZJ-1 strain of newcastle disease virus, they were serially passage to 20th on specific-pathogen-free chicken embryo fibroblast （CEF） monolayers. The purity of the rFPVs was detected by blue plaque assay and results showed that the rFPVs were stable, with 100 % plaque blue. The F genes of passages 0, 10 and 20 were amplified by PCR and sequenced, and there were not any changes on the nucleotide and amino acid. Subsequently expressions of F genes in CEF infected with the rFPVs were identified too by indirect immunofluoresence assay. All the results showed that the two rFPVs have good genetic stability for at least 20 passages.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2006年第4期405-407,共3页
Chinese Journal of Preventive Veterinary Medicine
基金
国家十五"863"计划(课题编号2001AA213041)
科技部农业科技成果转化资金项目(合同编号OIEFN213201078)资助
关键词
新城疫病毒
F基因
重组鸡痘病毒
遗传稳定性
newcastle disease virus
fusion protein gene
recombinant fowlpox virus
genetic stability
