摘要
人与小鼠和牛在正常胚胎植入前发育过程中基因活化的研究已经取得了长足的进展 ,但是还没有对同期克隆胚胎相关研究的报道 .利用单个家兔植入前移核重构胚胎成功地构建了MⅡ卵母细胞及发育至 4 、8 细胞期的胚胎和囊胚的特异性cDNA文库 .并用 β肌动蛋白和LAPTM4α证实这类文库是可靠的 .以 8 细胞期移核重构胚cDNA文库为例 ,随机挑取克隆进行测序分析 :其中 2 3的基因EST片段可以在GenBank或EST库中找到同源序列 ,约 1 3的EST片段属于未知的新片段 ,表明这是一类重要的新兴基因资源库 (期特异性EST库 ) .这种利用单胚胎构建cDNA文库的方法 ,解决了胚胎研究材料受限的问题 ,在时间上更加精确 ,更符合胚胎发育的规律 ,也能够更加准确地反映出一些克隆胚胎的异常表型 ,是研究早期胚胎发育基因表达以及克隆胚胎再程序化基因表达的一种有效手段 .
The gene activity during rabbit preimplantation cloned embryos development has not been reported,despite great progress having been made recently about normal preimplantation development in human,mouse and bovine.The stage\|specific cDNA libraries were constructed from single four\|cell,eight\|cell and blastocyte embryos,respectively.They have average complexities of 1 0×10\+6 independent clones.From the single eight\|cell cDNA libraries,13 clones were randomly selected to sequence.2/3 of the sequence were homologous to known transcripts or expressed sequence tags (ESTs) from human,mouse,and bovine.Novel sequences were detected at a frequency of 1/3.Reverse Northern blot analyses of specific and general genes (LAPTM4α and β\|actin) proved the credibility of these libraries.These cDNA libraries are a valuable resource for the isolation of clones representing genes and nuclear reprogramming active at these early development stage.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2004年第2期162-165,共4页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家重大基础研究发展规划 (973 )资助(No .2 0 0 0 0 1610 7)~~
关键词
基因活化
克隆
胚胎
CDNA文库
兔
rabbit preimplantation development,embryonic expression libraries,reprogramming related gene,ESTs,nuclear transfer
