纽荷尔脐橙贮藏枯水相关ESTs分离与初步分析

Isolation of Granulation-related ESTs in Newhall Navel Orange Fruit Storage and Primary Analyses of the Sequences

为分析柑橘果实贮藏过程中枯水相关特异基因的表达情况,以纽荷尔脐橙(Citrus Sinensis Osbeck)采摘后贮藏至略见枯水果实的果肉为测验方(Tester),以刚采摘果实的果肉为驱动方(Driver),采用抑制性差减杂交(Suppression Subtractive Hybridization,SSH)技术成功构建了贮藏果实差减cDNA文库,共获得656个阳性克隆。随机挑取克隆进行菌液PCR分析,插入片段长度主要集中在100～1000bp。成功对213个克隆进行测序,获得有效序列143条,经序列分析,共得到53条uniESTs序列。在NCBI基因库中对这53个uniESTs进行BLAST分析,比对结果参照MIPS的分类标准,按功能分为14类,其中参与代谢、能量及蛋白合成的ESTs最多,分别占到了全部ESTs的17%、11%和13%。

In order to analyze the differentially expressed genes in the post-harvested citrus fruit,a subtracted cDNA library was constructed by suppression subtractive hybridization（SSH）between the fruit pulp of post-storage（experimental group,the Tester）and that of pre-storage（control group,the Driver）.The library included about 656 clones which were 100-1 000 bp in size,among which 213 clones were selected randomly and sequenced.Totally 143 useful ESTs were acquired,and 53 uniESTs were finally obtained.Sequence similarity searches via BLASTx and BLASTn on NCBI showed that the ESTs were sorted into 14 functional categories according to the Munich Information Center for Protein Sequences（MIPS）.The ESTs involved in metabolism,energy,protein synthesis accounted for 17%,11% and 13% of the total ESTs,respectively.