›› 2010, Vol. 37 ›› Issue (10): 130-134.

• 遗传繁育 • 上一篇    下一篇

发育潜能不同的猪孤雌胚胎差异表达基因的筛选

陆芳1,2, 罗学明2, 龙川2, 王希彪1, 潘登科2   

  1. (1. 东北农业大学动物科技学院, 哈尔滨 150030;2. 中国农业科学院北京畜牧兽医研究所,农业部畜禽遗传资源与利用重点实验室, 北京 100193)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-10-20 发布日期:2010-10-20
  • 通讯作者: 潘登科

Screening of Differentially Expressed Genes in Pig Parthenogenetic Embryos with Different Developmental Competence

LU Fang1,2,LUO Xue-ming2,LONG Chuan2,WANG Xi-biao1,PAN Deng-ke2   

  1. (1.College of Animal Science and Technology, Northeast Agricultural University,Harbin 150030,China;2.The Key Laboratory of Farm Animal Genetic Resources and Utilization of Ministry of Agriculture,Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-10-20 Published:2010-10-20
  • Contact: PAN Deng-ke

摘要: 本试验以猪孤雌胚胎为材料,根据初次卵裂时间的早晚划分出发育能力不同的2组胚胎,比较其母源mRNAs丰度。利用mRNA差异显示技术(mRNA differential display reverse transcription PCR,DDRT-PCR)筛选出2组胚胎中表达丰度不同的母源mRNAs,并经反Northern blotting验证,结果发现,有8条差异表达的阳性cDNA片段(E1~E4; L1~L4),克隆、测序后与GenBank数据库进行同源性比对分析,其中6条(E1、E2、E4、L1、L2、L4)与已知ESTs有较高的同源性,E3与人CPEB2的同源性为97%,L3与人hnRNPA1的同源性为89%。因此,CPEB2和hnRNPA1是影响胚胎发育能力的重要候选基因。

关键词: 猪; 孤雌胚胎; 初次卵裂时间; 母源mRNA; 胚胎质量; DDTR-PCR

Abstract: The study obtained developmental competent and incompetent two-cell stage pig parthenogenetic embryos based on different first cleavage time and the level of maternal mRNAs was compared. Differential display reverse transcription polymerase chain reaction was carried out on maternal mRNAs isolated from these embryos. Eight fragments (E1 to E4, L1 to L4) differentially expressed between the two groups by reverse Northern blotting. After cloning and sequencing, eight homologue sequences were retrieved from NCBI database: six ESTs, E3 had high homology (the homology was 97%) with human CPEB2 and L3 had high homology (the homology was 89%) with hnRNPA1. CPEB2 and hnRNPA1 might play major roles in the developmental competence of embryos.

Key words: pig; parthenogenetic embryo; first cleavage time; maternal mRNA; embryo quality; DDRT-PCR

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