作　　者： ; ; ; ; ; ; ; ;

机构地区： 南京医科大学基础医学院生物化学与分子生物学系

出　　处： 《南京医科大学学报（自然科学版）》 2013年第4期426-431,共6页

摘　　要： 目的:探讨黑色素瘤抑制蛋白(melanoma inhibitory activity,MIA)在非洲爪蟾早期胚胎发育中的作用。方法:根据热带爪蟾MIA序列在非洲爪蟾EST数据库中找到MIA序列,利用RT-PCR扩增全长序列并克隆至pCS2+,用于测序。利用RT-PCR检测胚胎发育各阶段MIA mRNA的表达。通过显微注射特异的反义寡核苷酸morpholino(MO)敲降MIA;利用整胚原位杂交检测胚层标志性基因的表达。结果:克隆出非洲爪蟾MIA全长序列。在胚胎发育过程中MIA从受精卵开始持续表达。敲降MIA导致胚胎胚孔闭合延缓,中胚层标志基因xbra表达明显受抑,而内胚层标志性基因sox17α表达无明显改变。结论:MIA表达于非洲爪蟾胚胎发育各阶段,敲降MIA抑制中胚层的形成。 Objective：To investigate the effect of melanoma inhibitory activity （MIA） on mesoderm formation during the embryogenesis of Xenopus lavies. Methods：The MIA sequence of Xenopus lavies was verified in EST database, and then cloned into pCS2＋ for sequencing. The expression of MIA was detected during embryo development by RT-PCR analysis. The micro-injection of morpholino （MO） was used to knockdown MIA. The expression of marker genes was detected during germ layer formation by whole mount in situ hybridization. Results：The MIA sequence of Xenopus lavies was precisely cloned. The expression of MIA was detected at all the stages during embryogenesis. The knockdown of MIA by MO injection caused the delayed blastopore closure, inhibited the expression of mesodermal marker gene xbra compared to normal embryos. Conclusion：MIA is expressed during embryogenesis of Xenopus lavies, and MIA could play an important role in the mesoderm formation.

关 键 词： 黑色素瘤抑制蛋白 非洲爪蟾 胚胎发育 中胚层

领　　域： [生物学]