FANG Zi-yi,SONG Xiao-xi,ZHAO Shi-ping,et al.Cloning and Sequence Analysis of Aquaporins Gene in Chimonanthus praecox cv.Luteus[J].Northern Horticulture,2013,37(21):99-104.
素心蜡梅水孔蛋白基因的克隆与序列分析
- Title:
- Cloning and Sequence Analysis of Aquaporins Gene in Chimonanthus praecox cv.Luteus
- 文章编号:
- 1001-0009(2013)21-0099-06
- 分类号:
- S 685.99
- 文献标志码:
- A
- 摘要:
- 以素心蜡梅为试材，根据已建立的素心蜡梅转录组数据库已知序列设计特异引物，采用PCR技术，克隆并分析了素心蜡梅盛花期花被片水孔蛋白基因序列和氨基酸结构特征，对得到的基因和数据库中的cDNA进行比对，并通过多种生物信息学方法对其编码序列、蛋白结构和进化关系进行了分析预测。结果表明：该试验成功克隆得到长2 264 bp的水孔蛋白基因，命名为CpAQP3。经序列比对，蜡梅CpAQP3基因含5’非编码区50 bp，3’非编码区165 bp，开放阅读框长2 049 bp，编码248个氨基酸。该基因含2个内含子，长度分别为708 bp和594 bp，3个外显子，长度分别为127、250、370 bp。预测该蛋白有多个跨膜螺旋结构，具较强疏水性。该基因属于MIP家族中的TIP亚族，具有家族典型的保守结构域，预测其结构与运输水分具有显著关系。该试验首次从素心蜡梅中克隆得到水孔蛋白基因序列，与已知从磬口腊梅中获得的水孔蛋白序列有明显不同。
- Abstract:
- Taking Chimonanthus praecox cv.Luteus as material，a pair of primers were designed according to the gene sequence in the transcriptome database created，the gene encoding aquaporins in sepals of Chimonanthus praecox at peak flowering stage was cloned by PCR and its molecular structure of amino acid was analyzed.Based on sequence alignment in cDNA database and using bioinformatics，the coding sequence，protein structure and evolutionary relationships were analyzed.The results showed that a 2 264 bp gene encoding aquaporins was amplified successfully from genomic DNA of petals of Chimonanthus praecox at peak flowering stage，being named CpAQP3.The gene CpAQP3 consisted a 5′untranslated region (UTR) of 50 bp，a single long open reading frame (ORF) of 2 049 bp，and a 3′UTR of 165 bp.The ORF encoded 284 amino acids.This gene include two introns of 708 bp and 594 bp and three exons of 127 bp，250 bp and 370 bp respectively.An aquaporin gene CpAQP3 was firstly cloned from Chimonanthus praecox.The CaAQP gene was a member of the TIP subgroup in MIP protein family and had the typical function region.The CpAQP3 gene may play an important regulating water transportation role.There were significant differences in sequential structure between this gene and the other known aquaporin gene from Xinkou Chimonanthus praecox.
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备注/Memo
第一作者简介：方子义（1987-），男，硕士研究生，研究方向为园林植物种植资源创新及利用。