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Hereditas
2015年
11期
1160-1166
,共7页
减数分裂%SC%C(2)M%酵母双杂交%蛋白相互作用
減數分裂%SC%C(2)M%酵母雙雜交%蛋白相互作用
감수분렬%SC%C(2)M%효모쌍잡교%단백상호작용
meiosis%SC%C(2)M%yeast two-hybrid system%protein interaction
同源染色体联会时形成的联会复合体(Synaptonemal complex, SC)是由减数分裂前期Ⅰ多种蛋白质聚集而成的超级复合结构。生殖细胞特异性的核蛋白C(2)M(Crossover suppressor on 2 of Manheim)在染色体上高度聚集可以诱导SC的形成。本文采用酵母双杂交方法,利用C(2)M的诱饵表达载体筛选果蝇cDNA文库,共发现40个可能与C(2)M相互作用的蛋白,包括多种DNA及组蛋白结合蛋白、ATPase、转录调节因子。从筛选的结果中,选取wech和Psf1基因构建了转基因果蝇,并在生殖细胞中进行了基因沉默,结果显示联会复合体的消失受到延迟。上述结果表明Wech和Psf1蛋白可能与C(2)M形成复合物,共同参与联会复合体的形成或其稳定性的维持。
同源染色體聯會時形成的聯會複閤體(Synaptonemal complex, SC)是由減數分裂前期Ⅰ多種蛋白質聚集而成的超級複閤結構。生殖細胞特異性的覈蛋白C(2)M(Crossover suppressor on 2 of Manheim)在染色體上高度聚集可以誘導SC的形成。本文採用酵母雙雜交方法,利用C(2)M的誘餌錶達載體篩選果蠅cDNA文庫,共髮現40箇可能與C(2)M相互作用的蛋白,包括多種DNA及組蛋白結閤蛋白、ATPase、轉錄調節因子。從篩選的結果中,選取wech和Psf1基因構建瞭轉基因果蠅,併在生殖細胞中進行瞭基因沉默,結果顯示聯會複閤體的消失受到延遲。上述結果錶明Wech和Psf1蛋白可能與C(2)M形成複閤物,共同參與聯會複閤體的形成或其穩定性的維持。
동원염색체련회시형성적련회복합체(Synaptonemal complex, SC)시유감수분렬전기Ⅰ다충단백질취집이성적초급복합결구。생식세포특이성적핵단백C(2)M(Crossover suppressor on 2 of Manheim)재염색체상고도취집가이유도SC적형성。본문채용효모쌍잡교방법,이용C(2)M적유이표체재체사선과승cDNA문고,공발현40개가능여C(2)M상호작용적단백,포괄다충DNA급조단백결합단백、ATPase、전록조절인자。종사선적결과중,선취wech화Psf1기인구건료전기인과승,병재생식세포중진행료기인침묵,결과현시련회복합체적소실수도연지。상술결과표명Wech화Psf1단백가능여C(2)M형성복합물,공동삼여련회복합체적형성혹기은정성적유지。
The synaptonemal complex (SC) is a huge structure which assembles between the homologous chro-mosomes during meiotic prophase I.Drosophila germ cell-specific nucleoprotein C(2)M clustering at chromo-somes can induce SC formation. To further study the molecular function and mechanism of C(2)M in meiosis, we constructed a bait vector for C(2)M and used the yeast two-hybrid system to identify C(2)M interacting proteins. Forty interacting proteins were obtained, including many DNA and histone binding proteins, ATP synthases and tran-scription factors. Gene silencing assays inDrosophila showed that two genes,wech andPsf1, may delay the disap-pearance of SC. These results indicate that Wech and Psf1 may form a complex with C(2)M to participate in the for-mation or stabilization of the SC complex.