茶树花粉特异蛋白基因CsPSP1的分离及序列分析  

Isolation and Characterization of CsPSP1,a Gene Encoding Pollen Specific Protein in Tea Plant(Camellia sinensis)

作　　者：余梅[1] 江昌俊[1] 叶爱华[1] 王朝霞[2] 朱林[3] 

机构地区：[1]安徽农业大学茶叶生物化学和生物技术教育部,农业部重点实验室,安徽合肥230036 [2]合肥学院生物系,安徽合肥230061 [3]安徽农业大学资源与环境学院,安徽合肥230036

出　　处：《激光生物学报》2008年第2期206-212,共7页Acta Laser Biology Sinica

基　　金：安徽省教育厅自然科学研究计划重点项目(2004KJ137ZD)

摘　　要：利用cDNA-AFLP技术比较了茶树[Camellia sinensis(L.)O.Kuntze cv.Wulong]花蕾发育早期和晚期的基因表达,结果表明存在明显差异。以E12和M20为引物对在晚期发育花蕾中筛选出一条281 bp特异表达的差异条带TDF53(transcipt-derived-fragment,TDF)。RT-PCR分析表明该片段只在晚期发育花蕾中特异表达。用RACE方法延伸其末端序列,克隆并测序获得全长cDNA序列(GenBank登录号:DQ887753)。该基因全长2079 bp,开放阅读框1701 bp,编码567个氨基酸,其分子量为63 kDa。序列和结构的同源性分析表明:该基因编码的氨基酸序列与烟草、油菜的花粉特异蛋白等同源性较高,由此推定,该基因为编码茶树花粉特异蛋白的基因,并将分离到的花粉特异蛋白基因命名为CsPSP1。The eDNA-amplified fragment length polymorphism approach was used to identify genes expressed differentially during early and late flower bud development in tea plant （ Camellia sinensis）. The result showed that there was evident differential expression in flower buds between the early and late stages. A transeript derived fragment, TDF53, was obtained via selective amplification with E12/M20 primer pair. TDF53 was specifically expressed only in the flower buds at the late stages by RT-PCR. The complete eDNA was recovered by rapid amplification of eDNA ends, then cloned and sequenced. It was found that the full-length eDNA of 2 079 bp （ GenBank accession number DQ887753 ） included a 1 701 bp open reading frame predicting a 567-amino-acid polypeptlde of 63 kDa. Analysis of the nueleotlde sequence and deduced amino acid sequence revealed a high homology with pollen specific proteins from tobacco and Brassica napus. Thus, we propose that this eDNA encodes the first pollen specific protein described in tea plant, designated CsPSP1.

关 键 词：花粉特异蛋白 CDNA 茶树 花蕾 差异表达 

分 类 号：Q78[生物学—分子生物学]