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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 expression of dry-related genes in citrus fruits during storage, Citrus Sinensis Osbeck was used to pick the flesh which was slightly seeded in dry fruits as the test organ (Tester) As a driver, a total of 656 positive clones were successfully constructed by Suppression Subtractive Hybridization (SSH) technique. Randomly picked clones for bacterial PCR analysis, the length of the insert is mainly concentrated in the 100 ~ 1000bp. 213 clones were successfully sequenced and 143 valid sequences were obtained. A total of 53 uniESTs sequences were obtained by sequence analysis. The 53 uniESTs were analyzed by BLAST in the NCBI GenBank. The results of the alignment were classified into 14 categories according to the classification criteria of MIPS. Among them, ESTs involved in metabolism, energy and protein synthesis accounted for 17% , 11% and 13% respectively.