琯溪蜜柚汁胞Beta-tubulin cDNA的克隆及序列分析  

Cloning and sequence analysis of Beta-tubulin cDNA of juice sac in Citrus grandis (L.) Osbeck

作　　者：钟凤林[1,2] 郭志雄[1] 林琳[1] 马传营[1,2] 潘东明[2] 

机构地区：[1]福建农林大学园艺学院 [2]福建农林大学园艺产品贮藏保鲜研究所,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2010年第3期241-245,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：国家科技支撑计划资助项目(2007BAD07B01)

摘　　要：以琯溪蜜柚汁胞为材料,采用cDNA克隆方法进行Beta-tubulin cDNA的克隆和表达.结果表明:琯溪蜜柚果肉Beta-tubulin cDNA全长1636 bp,有完整的阅读框,编码444个氨基酸;5′非翻译区60 bp,3′非翻译区243 bp(不包含终止密码子TAA),其中包括1个多聚腺苷酸化信号AATAAA以及1个含21个腺苷酸的poly(A)尾.由琯溪蜜柚果肉Beta-tubulin cDNA推导的氨基酸序列与陆地棉(Gossypium hirsutum)、大豆(Glycine max)、杨属植物(Populus)等的同源性较高,分别为86%、85%、84%等.通过构建重组质粒p28-PTU,并转化至Trans Rosetta(DE3)中,在诱导温度为37℃、IPTG浓度为1 mm、诱导时间为4 h的条件下,其在大肠杆菌宿主中获得了表达.The pummelo（Citrus maxima（L.） Osbeck） juice sac Beta-tubulin cDNA was cloned,sequenced and analyzed.The results showed that the complete cDNA was 1636 bp in length,including 60 bp 5′ untranslated region,243 bp 3′ untranslated region which contained a polyadenylation signal AATAAA and a 21 bp polyadenylation tail.The coding region encoded a peptide of 444 amino acid residues.The deduced amino acid sequence deduced by the Beta-tubulin sequences of juice sac in Citrus grandis（L.） had higher homology with upland cotton（Gossypium hirsutum）,fig（Ficus carica） and poplar（Populus）,which were 86%,85% and 84%,respectively.The coding region of the cDNA was ligated into pET-28a vector to build a recombinant vector p28-PTU,which was expressed in Escherichia Coli Trans Rosetta（DE3） strain when induced with 1 mm IPTG and grown for 4 hours at 37 ℃.

关 键 词：琯溪蜜柚 BETA-TUBULIN CDNA 序列分析 同源性分析 

分 类 号：S666.3[农业科学—果树学]