【作者】 佘文琴； 潘东明； 陈桂信； 郭志雄；

【Author】 SHE wen-qin,PAN dong-ming~*,CHEN gui-xin,and GUO zhi-xiong College of Horticulture,Fujian Agricultural and Forestry University,Fuzhou 350002 China

【机构】 福建农林大学园艺学院；

【摘要】 本研究以琯溪蜜柚果肉汁胞为材料,运用RT-PCR的方法,得到了琯溪蜜柚果实汁胞粒化过程中未粒化汁胞的差异片断,通过Blast分析发现该片段序列与从发育中桃果实分离得到的cDNA有高度的同源性。在此基础上,根据汁胞未粒化基因差异cDNA片段Miyoul设计引物,采用RACE技术克隆了该片段5’端全长序列,将获得的5’端cDNA与Miyoul差异片段拼接,拼接后得到Miyoul全长序列,其大小为1152 bp,包含1个819 bp的的开放阅读框架,编码272个氨基酸。其中,5′UTR为77 bp,3′UTR为244 bp,3′UTR区域还含有poly(A)尾。登录号为F.J866624。进一步的Blast分析显示,该cDNA全长序列与20S蛋白质酶体的β亚基5高度同源。其中,与鹰嘴豆的20S蛋白质酶体的β亚基5同源性较高,为84.9%,与水稻、小麦的同源性分别为76.1%和74.6%。20S蛋白质酶体与蛋白质的降解有关。更多还原

【Abstract】 Guanxi-miyou fruit being taken as experimental material in this study,CDNA difference fragment of non-granulated juicy sac was isolated in the process of granulation of Guangxi-miyou by reverse transcription-polymerase chain reaction(RT-PCR).It was found that the Guanxi-miyou CDNA fragment was significantly homologous to developing pear fruit’s by Blast analysis.According to Miyoul cDNA fragments designed primers of juice sacs normal gene differential cDNA fragments,5′full-length sequence was cloned by RACE technique and jointed with Miyoul fragment,and then cDNA full length with the size of 1152 bp was gained which included a open reading frame of 819 bp and encoding 272 amino acid.Its accession number of GenBank database was FJ866624.Thereamong the size of 5′UTR was 77bp,that of 3′UTR was 244 bp and 3′UTR region contained poly(A) tail.The further blast analysis showed,this cDNA had high sequence identity withβ5 subunit of 20s proteasome.It had the highest homology with that of chickpea (84.9%),and next with that of rice(76.1%) and with that of wheat(74.6%).Proteasome is a kind of proteolytic enzyme compounds exiting in cell cytoplasm and nucleus and plays a important role in degrading the intracellular protein cell protein and maintaining cell normal metabolism,therefore 20s proteasome plays a role in sac growth of‘Guanxi-miyou’ fruit.更多还原