摘要
研究旨在获得PCV2a和PCV2b的拯救毒株。对筛选得到的PCV2a和PCV2b感染性克隆分别进行酶切,使酶切得到的全长基因组DNA自身环化并分别转染PK15细胞。经IFA和PCR验证,确认成功拯救出两亚型PCV2病毒。病毒传至第9代时对PCV2a和PCV2b的毒价分别进行测定,结果显示PCV2a的毒价为103.5 TCID50/mL,PCV2b的毒价为104.6 TCID50/mL。本试验为分型特异性序列与病毒致病力相关性的研究打下了基础。
The study was designed to get the rescued strains of PCV2a and PCV2b. The screened PCV2a and PCV2b infectious clones were digested, then the genomic DNA was cyclized and transfected in- to PK15 cells. IFA and PCR indicated that two subtypes of PCV2 were rescued successfully. Virus titers in cell cultures for the 9th passage were determined by a microtitration infectivity assay and recorded as TCID50/mL. PCV2a was 103^3.5 TCID50/mL,and PCV2b was 104.6 TCID50/mL. This experiment laid the foundation for the study of the correlation between type-specific sequence and virus virulence.
出处
《家畜生态学报》
北大核心
2012年第6期67-71,共5页
Journal of Domestic Animal Ecology
基金
山东省自然科学基金资助项目(2010ZRC02031)
国家成果转化项目(2011GB2C600011,2010GB2C600259)
关键词
猪圆环病毒2型
分型特异性序列
自身环化
体外拯救
porcine circovirus type 2 （PCV2）
type-specific sequence
the ligation DNA of PCV2
char- acterizing in vitro
