摘要
[目的]克隆枳早期结瘤素样基因PtBCP1并对其序列进行分析。[方法]以枳消减文库中的C28 EST为种子序列进行电子克隆,据此设计引物进行PCR,以获得枳早期结瘤素样基因PtBCP1的全长cDNA和DNA序列,并对获得的序列进行生物信息学分析。[结果]PtBCP1基因由2个外显子和1个内含子组成,在枳幼苗根中的表达量是叶中的140倍,该基因编码的蛋白含131个氨基酸残基,预测分子量为14.0 kD,理论等电点为8.75,具有N端信号肽和PCLD(PFMD:PF02298)功能域,但无完整的铜离子结合位点,属于早期结瘤素样蛋白。[结论]为进一步研究PtBCP1基因的生物学功能奠定了基础。
[Objective] The aim was to clone and analyze the sequence of phytocyanin-related early nodulin-like gene（PtBCP1） from Poncirus trifoliata.[Method] The electronic cloning was done by using the C28 EST of Poncirus trifoliata subtractive library as seed sequences and the RT-PCR primers were designed by that.Then the PtBCP1 cDNA and DNA of Poncirus trifoliate were cloned by PCR and the sequences were analyzed by bioinformatics methods.[Result] The PtBCP1 gene included two exons and one intron,and was 140 times of expression in the seeding root of Poncirus trifoliata than that of in leaves.The gene encoded a polypeptide of 131 amino acids with a 14 kD molecular weight and 8.75 isoelectric point.The PtBCP1 protein was a early nodulin-like protein,with a signal peptide in the N-terminal and a plastocyanin-like domain in the C-terminal,and without the complete copper-binding sites.[Conclusion] It prepared the ground for the further research of PtBCP1 gene biological function.
出处
《安徽农业科学》
CAS
2012年第33期16102-16106,共5页
Journal of Anhui Agricultural Sciences
基金
长江学者和创新团队发展计划项目(PCSIRT)
柑桔产业技术体系(CARS-27)农业部公益性行业(农业)科研专项(2010 03067-02-4)
重庆市自然科学基金计划项目(cstc2011jjA811)
关键词
枳
早期结瘤素样基因
克隆
Poncirus trifoliata
Early nodulin-like gene
Clone
