表达新城疫病毒F基因重组鸡痘病毒的遗传稳定性

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，rFPV Lp-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rFPV LP-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：艮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。

Genetic stability of recombinant fowlpox virus vaccine expressing fusion protein gene of newcastle disease virus

In order to evaluate the genetic stability of two recombinant fowlpox viruses(rFPV282E4-FSrFPVLP-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.