用cDNA芯片技术筛选中华绒鳌蟹卵巢发育相关基因  

Screening the Differentially Expressed Genes Related to the Ovarian Development of Chinese Mitten Crab(Eriocheir japonica sinensis) Using cDNA Macroarray

作　　者：马长艳[1,2] 郭豫杰[1] 周开亚[1] 

机构地区：[1]南京师范大学生命科学学院,江苏省生物多样性和生物技术重点实验室,江苏南京210046 [2]南京医科大学发育遗传学系,江苏南京210029

出　　处：《南京师大学报（自然科学版）》2010年第3期76-80,共5页Journal of Nanjing Normal University(Natural Science Edition)

基　　金：国家自然科学基金(30870286);国家自然科学基金重点项目(30630010)

摘　　要：为了克隆中华绒螯蟹卵巢发育相关基因,我们构建了中华绒螯蟹卵巢差减cDNA文库.本研究用cDNA芯片技术对中华绒螯蟹卵巢差减cDNA文库进行了初步筛选,并从cDNA芯片筛选的正向差减文库中随机选取50个克隆进行序列测定.结果共获得2倍以上差异表达克隆167个,其中Ⅲ期高表达克隆104个,Ⅱ期高表达克隆63个.正向差减文库中共获得7个独立的EST(expressed sequence tag,EST).同源性分析结果表明,这些EST皆为新的EST,dbEST登录号分别为:CA591892、CA591893、CA591894、CA591895、CA591896、CA591897和CA591898.本研究结果为进一步克隆中华绒螯蟹卵巢发育相关基因的全长cDNA序列并研究其功能,阐明中华绒螯蟹卵巢发育的分子机理奠定了重要的前期工作基础.To clone genes which are related to the ovarian development of Chinese mitten crab,the subtracted libraries have previously been constructed.In the present study,the cDNA macroarray was prepared to screen the differentially expressed genes and fifty clones identified from the forward subtracted cDNA library were selected randomly and sequenced.In total,167 clones whose signal difference was〉2 folds were obtained.Of the differentially expressed clones,104 clones were highly expressed in the ovary at stage Ⅲ,63 clones were highly expressed in the ovary at stage Ⅱ.Moreover,seven independent ESTs were obtained from the forward subtracted cDNA library.Homology analysis showed that there aren＇t sequences significantly matching to these ESTs.So they were deduced to be novel ESTs,and all of them were deposited in dbEST.The accession numbers are： CA591892、CA591893、CA591894、CA591895、CA591896、CA591897 and CA591898,respectively.

关 键 词：中华绒螯蟹 卵巢 抑制性差减杂交 CDNA芯片 表达序列标签 

分 类 号：Q75[生物学—分子生物学]