【作者】 钟凤林； 林琳； 郭志雄； 马传营； 潘东明；

【Author】 Zhong Feng-lin~(1,2),Guo Zhi-xiong~1,Lin Lin~1,Ma Chuan-ying~(1,2),and Pan Dong-ming~(2*) (1 College of Horticulture,Fujian Agriculture and Forestry University,Fuzhou 350002;2 Institute of Storage Science and Technology of Horticultural Products,Fujian Agriculture and Forestry University,Fuzhou 350002)

【机构】 福建农林大学园艺学院； 福建农林大学园艺产品贮藏保鲜研究所；

【摘要】 本研究以琯溪蜜柚汁胞为材料,运用cDNA克隆的方法,克隆得到了琯溪蜜柚果肉过氧化物酶的cDNA的全长,并对得到的序列进行了一些分析,从分子生物学角度对粒化的研究做了初步的探索。结果表明:琯溪蜜柚果肉Beta-tubulin cDNA全长1636bp,有完整的阅读框,编码444个氨基酸。另外5′非翻译区60bp,3′非翻译区243bp(不包含终止密码子TAA),其中包括一个多聚腺苷酸化信号AATAAA,以及一个含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)等同源性较高,分别为:86%,85%,84%等。通过构建重组质粒p28-PTU,转化至Trans Rosetta(DE3)中,当诱导温度为37℃,IPTG浓度为1mM,诱导时间为4h时,其在大肠杆菌宿主中获得了表达。更多还原

【Abstract】 The pummelo(Citrus maxima(L.) Osbeck) juice sac Beta-tubulin cDNA was cloned, sequenced and analysised.The results showed that the complete cDNA was 1636bp in length,including 60bp of 5′untranslated region,243bp of 3′untranslated region which contained a polyadenylation signal,AATAAA, and a 21bp polyadenylation tail.The coding region encoded a peptide of 444 amino acid residues.The deduced amino acid sequence had a significant homology to the Beta-tubulin sequences from upland cotton (Gossypium hirsutum),fig(Ficus carica) and poplar(Populus),were 86%,85%,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℃.更多还原