朊蛋白基因在奶牛生殖系统不同组织中的表达量测定  

Prion gene expression in different tissues of cow reproductive system

作　　者：张太翔[1] 杨建民[1] 周向梅[1] 尹晓敏[1] 赵德明[1] 

机构地区：[1]中国农业大学国家动物海绵状脑病实验室,北京100094

出　　处：《中国预防兽医学报》2007年第4期263-267,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然基金(No.30571399;30500371);教育部博士点基金(No.20050019031)资助项目

摘　　要：为了研究疯牛病能够垂直传播的理论基础,我们采用PrP基因做目的基因,GAPDH基因做内参照的相对荧光定量RT-PCR方法,对目的基因和内参基因分别构建标准重组质粒,制备标准曲线,利用标准曲线对采集雌雄奶牛生殖系统的12个不同组织PrP基因表达进行定量。试验结果表明雌雄奶牛的生殖系统各部分组织都有PrP基因表达,其中雄性奶牛的睾丸、附睾和输精管呈现高的表达量,分别为1.16±0.0010、1.81±0.0056、0.68±0.0038;前列腺、精囊腺和尿道球腺呈现中等表达量,分别为0.47±0.0018、0.25±0.0007、0.28±0.0007;阴茎呈现低的表达量为0.18±0.0017;雌性奶牛的子宫、子宫阜呈现高的表达量,分别为2.53±0.0008、1.75±0.0098;卵巢和输卵管呈现中等表达量,分别为1.55±0.0038、1.45±0.0064;阴道呈现低表达量为0.57±0.0036。本研究为阐释PrP基因在奶牛生殖系统的正常表达和正常的生理功能,以及为确定生殖系统在疯牛病垂直传播中的地位和其分子机理奠定了基础。To investigate the possibility of the maternal transmission of BSE, PrP mRNA expression in different tissues of cow reproductive system was quantified Using calibrated highly sensitive externally standardized real-time RT-PCR. Absolute quantification was achieved using a standard curve constructed for the target gene PrP and the housekeeping gene GAPDH. The results showed that PrP mRNA was present in all regions of the reproductive system of male and female cows. In bulls examined, the spermary, spermary appendix and spermaduct all showed high level of PrP mRNA expression （1.16±0.0010, 1.81 ± 0.0056, 0.68 ± 0.0038）; moderate level of the expression （0.47± 0.0018, 0.25 ± 0.0007, 0.28± 0.0007） were detected in the prostate, spermatic gland and urethral gland and low level of expression （0.18 ±0.0017） in the penis. In comparison, cows showed high level of PrP mRNA expression （2.53 ± 0.0008, 1.75 ± 0.0098） in the placenta and the uterus; moderate level of expression （1.55 ± 0.0038, 1.45± 0.0064） in the ovary and fallopian tube; and low level of expression （0.57 ± 0.0036） in vagina. The results arc useful in elucidating the normal PrP gene expression and its normal physiological functions, and to confirm the role of the reproductive system in the maternity transmission route of the BSE.

关 键 词：朊蛋白基因 奶牛生殖系统 MRNA表达 实时荧光定量RT-PCR 

分 类 号：S852.151[农业科学—基础兽医学]