摘要
应用抑制性差减杂交技术 ,已经获得了中华绒螯蟹卵巢发育过程中差异表达基因的部分cDNA序列。为了进一步获得基因的全长cDNA序列 ,运用SMART技术 ,成功构建了中华绒螯蟹卵巢 (Ⅲ期 )RACEcDNA文库。琼脂糖凝胶电泳结果表明 ,文库所含全长cDNA的长度主要集中在 5 0 0～ 2 0 0 0bp之间 ,RACEPCR结果表明 ,所用基因特异性引物与接头引物皆能扩增出产物 ,说明所构文库的质量较好 ,适于用RACE方法从中分离中华绒螯蟹卵巢发育相关基因的全长cDNA。
Using suppression subtractive hybridization (SSH) approach, partial cDNA sequences of the differentially expressed genes that are related to the development of the ovary of the mitten crab (Eriocheir japonica sinensis) were obtained. To get full-length cDNA sequences of these genes, a RACE cDNA library of the ovary during stage Ⅲ was constructed successfully with SMART technique. Agarose gel electrophoresis showed that the lengths of full-length cDNAs in this library were pooled mainly between 500 and 2?000 base pairs. By RACE PCR, amplified products were obtained with all the gene-specific primers and adaptor primers. This verified that the quality of the RACE cDNA library was high and was appropriate for cloning the full-length cDNAs of these genes.
出处
《动物学杂志》
CAS
CSCD
北大核心
2003年第2期14-16,共3页
Chinese Journal of Zoology
基金
教育部"2 11"工程建设项目
江苏省应用基础项目 (No .BJ95 10 9)
