HCLV在PK-15细胞中的滴度和12nt的变化

Change of a 12-nt and Titer of HCLV in PK-15 Cells

在PK-15细胞上连续传代获得的HCLV,从第1～25代病毒的测序结果一致,同源性为100%。从第26代开始在12-nt(ttttttctttttt)插入处出现变化,第26、27代在69位碱基C的左右各多插入了一个T;第28、29代在碱基C的右侧多插入了2个T;第30、31代在C的左侧多插入了6个T,碱基C也变成了T;第32、33代在C的右侧缺失了2个T,碱基C也缺失;第34代在C的左侧缺失了5个T,在C的右侧缺失了5个T,碱基C也缺失,并且在第86位由原来的A变成G,第92位由原来的T变成C。将HCLV1～34代病毒接种24孔板中长满单层的PK-15细胞,于接种后72h进行荧光抗体检查,比较不同代次HCLV在PK-15细胞上增殖的差异。结果发现,从28代开始病毒滴度有所下降。

HCLV strain was grown in Primary bovine testicular cells, from 1 to 34 serial passages were made continuously. The result of Sequence analysis showed The homologies of 3＇-UTR nucleotide sequence was 100 % from 1 to 25 serial passages. 12-nt（ttttttctttttt） insertion began to change from 26 serial passages, two T more than 1 serial passages in 69 place from 26 to 27 serial passages, Two T more than 1 serial passages in 69 place from 28 to 29 serial passages, six T more than 1 serial passages in 69 place from 30 to 31 serial passages, two T lack than 1 serial passages in 69 place from 32 to 33 serial passages, ten T more than 1 serial passages in 69 place 34 serial passages, A to G in 86 place, T to C in 92 place. HCLV strain from 1 to 34 serial passages cultured in PK-15 cells, immuno fluorescence assay in 72 h, Detection the difference of HCLV from 1 to 34 serial passages, detection indicated the titer of HCLV began to decrease from 28 serial passages.