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猪胎盘特异基因1(PLAC1)的克隆、组织表达及遗传方式研究
Cloning,Tissue Expression and Inheritance Patterns of Porcine Placenta-specific 1(PLAC1) Gene
【摘要】 旨在研究猪胎盘特异性基因PLAC1的表达规律,并对其基因结构、生物功能和遗传方式进行初步鉴定。本研究以妊娠26、50和95天的猪胎盘组织为材料,利用RACE-PCR及RT-PCR技术克隆猪PLAC1基因及其不同转录本的全长序列,同时通过原位杂交验证其在胎盘中的表达模式。并检测猪PLAC1基因的序列变异。结果表明,猪PLAC1基因存在3种不同的转录本,3种转录本的CDS区完全相同,长525bp,编码174个氨基酸;组织表达谱和原位杂交结果显示,PLAC1基因特异性表达于猪胎盘绒毛膜上皮细胞中,并在不同妊娠时期差异表达,妊娠50和95天PLAC1mRNA的表达显著高于妊娠26天(P<0.01)。猪群中PLAC1基因的SNP基因分型检测结果表明该基因遵循半合基因的遗传方式。本试验结果为研究猪PLAC1基因在胎盘发育过程中的生物功能奠定了基础。
【Abstract】 The objective of this study was to investigate the expression of porcine PLAC1 gene,and preliminarily identify its gene structure,biological function and inheritance patterns.The placenta tissue of pigs on 26,50 and 95 days of pregnancy were collected,the full-length of PLAC1 mRNA and its splicing variants were cloned by using the RACE-PCR(rapid-amplification of cDNA ends)and RT-qPCR(real-time quantitative PCR).Moreover,the expression patterns of PLAC1 gene in placenta of pigs was verified by in situ hybridization.In addition,the sequence variation of PLAC1 gene was detected.The results showed that there were 3 different transcripts of the PLAC1 gene in pigs,and the CDS regions of the 3 transcripts were identical,with a length of 525 bp,encoding 174 amino acids;The results of the expression profile and hybridization in situof PLAC1 gene indicated that the PLAC1 gene was specifically expressed in pig placentalchorionic epithelium,and the expression of PLAC1 mRNA was different during different stages of pregnancy.The expression of PLAC1 mRNA was significantly higher on 50 and 95 days of gestation in placenta of pigs than 26 days of gestation.The genotype of SNP on PLAC1 gene confirmed that it was a hemizygous gene,and followed the inheritance patterns of hemizygous gene.The experiment data provide a basis for exploring the biological function of porcine PLAC1 gene in placenta development.
- 【文献出处】 畜牧兽医学报 ,Chinese Journal of Animal and Veterinary Sciences , 编辑部邮箱 ,2019年01期
- 【分类号】S828
- 【下载频次】175