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国家自然科学基金(30960271)

作品数:4 被引量:12H指数:2
相关作者:刘淑英徐萌杰齐景伟韩硕更多>>
相关机构:内蒙古农业大学更多>>
发文基金:国家自然科学基金更多>>
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Expression of Endogenous Beta Retroviruses and Hyal-2 mRNA in Immune Organs of Fetuses and Lambs被引量:5
2012年
Endogenous beta retroviruses (enJSRV) are highly homologous with Jaagsiekte sheep retrovirus (exJSRV),this exogenous retrovirus is the aetiological agent of ovine pulmonary adenocarcinoma (OPA). The aim of this study was to clarify the function of enJSRV and the immunological mechanisms of its corresponding antibody, that is undetectable in JSRV-infected ovine serum. The expression of enJSRV envelope protein and Hyal-2 mRNA in immune organs and lungs of ovine fetuses and lambs were analyzed by Real-Time reverse transcription PCR and In Situ Hybridization using specific probes. In Situ Hybridization results indicated that the enJSRV envelope protein and Hyal-2 mRNA were expressed in thymus, spleen, mesenteric lymph nodes and lungs at different times, while no positive signals were detected in the negative controls. On the other hand, results from Real-Time reverse transcription PCR analysis showed that in 130d fetuses and 3d newborn lambs the enJSRV mRNA levels were much higher in organs associated with the immune system than that in lungs, especially in the thymus and spleen, but levels of Hyal-2 mRNA expression was not significantly different in all collected tissue. These results provided evidence from an immunology point of view to understand why the circulating antibodies against exJSRV are undetectable in JSRV-infected ovine, and will help to unravel the pathogenesis of JSRV-infected ovine.
Jing-wei QiXiao-li WuShu-ying LiuGui-fang Cao
关键词:新生羔羊MRNA表达水平MRNA水平
Identification of Sheep Endogenous Beta-Retroviruses with Uterus-Specific Expression in the Pregnant Mongolian Ewe被引量:6
2013年
The sheep genome harbours approximately 20 copies of endogenous beta-retroviruses(enJSRVs),and circumstantial evidence suggests that enJSRVs might play a role in mammalian reproduction,particularly placental morphogenesis.This study was aimed to assess the expression of mRNAs of an enJSRV and its receptor,HYAL2,in the uterus and conceptuses of Mongolian ewes throughout gestation,using real-time reverse transcription polymerase chain reaction and in situ hybridization analysis.The results showed that enJSRV and HYAL2 mRNAs were found to be expressed throughout gestation in the endometrium,chorion,placenta,and conceptus.The enJSRV mRNA was most abundant in the placenta on day 90 of pregnancy,in the endometrium on day 30 and 50,and in the chorion on day 70 and 110.However,HYAL2 mRNA was most abundant in the endometrium on day 30.These differences were all significantly different from each other(P<0.01).In situ hybridization showed that enJSRV and HYAL2 mRNAs were specifically expressed in endometrial luminal epithelium and glandular epithelium,trophoblastic giant binucleated cells(BNCs),endometrial caruncles,placental cotyledons,stroma,trophectoderm,as well as multinucleated syncytia of the placenta and blood vessel endothelial cells.Collectively,little is known about the molecular mechanisms by which trophoblastic differentiation and multinucleated syncytia formation are regulated by enJSRVs.However,the temporal and spatial distributions of enJSRV expression in the uterus and conceptus indicate that differentiation of BNCs and the formation of a multinucleated syncytiotrophoblast involve enJSRV and possibly its cellular receptor,HYAL2.Therefore,enJSRV and HYAL2 appear to play important roles in the female reproductive physiology in this breed of sheep.
QI Jing-weiXU Meng-jieLIU Shu-yingZHANG Yu-feiLIU YueZHANG Ya-kunCAO Gui-fang
关键词:内源性逆转录病毒特异表达基因腺上皮细胞
组织原位杂交和Taq Man实时荧光定量PCR检测enJSRV及其受体HYAL2在妊娠蒙古绵羊绒毛膜中的表达被引量:2
2012年
为了探究内源性绵羊肺腺瘤病毒(enJSRV)及其受体HYAL2在妊娠蒙古绵羊胚胎发育过程中的可能作用,本试验应用组织原位杂交技术和Taq Man实时荧光定量PCR对其在不同妊娠时期(30、50、70、90、110、130d)蒙古绵羊绒毛膜的分布定位和表达规律进行了研究。原位杂交结果显示,enJSRV及其受体HYAL2 mRNAs在妊娠30、50、130d蒙古绵羊绒毛膜组织的滋养层巨型双核细胞(BNC)、多核合胞体小半鞘翅(SP)和滋养外胚层(Tr)细胞中均有阳性信号表达;荧光定量PCR结果显示,在妊娠各时期绒毛膜组织中均有enJSRV及受体HYAL2的mRNA表达,通过统计学分析enJSRV mRNA的表达量于妊娠50d时最低,70d和110d相对较高,且差异都极显著(P<0.01),到130d时又降低到起始水平;受体HYAL2的表达于130d时表达量最低,90d相对较高,且差异极显著(P<0.01)。而enJSRV与其受体HYAL2 mRNA表达量之间无线性相关性。研究结果提示,enJSRV在绵羊胚胎发育过程中可能参与调节绒毛膜滋养层细胞的生长并影响胎盘的发育。
齐景伟徐萌杰刘淑英
关键词:内源性绵羊肺腺瘤病毒绒毛膜TAQ
绵羊肺腺瘤病毒内蒙株env基因的克隆及生物信息学分析
2010年
根据GenBank中绵羊肺腺瘤病毒(AF105220)env基因序列设计合成了1对引物。提取自然感染绵羊肺腺瘤病的羊肺组织总RNA,应用RT-PCR技术对env基因进行了扩增,将其回收后克隆入PMD19-T载体中并进行了序列测定。应用生物信息学分析技术并对env基因所编码的蛋白进行了结构预测。结果表明,env基因全长1 848bp,共编码615个氨基酸。在基因编码的TM区发现外源性exJSRV特异性的"YXXM"基序。测定序列与美国代表株(AF105220)的env基因序列比较核苷酸同源性为98.8%,推导出的氨基酸同源性为99.8%。通过protparam工具统计分析可知env基因所编码的蛋白为亲水蛋白。
韩硕刘淑英
关键词:绵羊肺腺瘤病毒ENV基因克隆
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