新疆农业科学2017，Vol.54Issue(10)：1765-1774,10.DOI:10.6048/j.issn.1001-4330.2017.10.001
哈密瓜果实转录因子CmCBF1的克隆及表达分析
Cloning and Expression Analysis of a CRT/DRE-binding Factor Gene CmCBF1 from Hami Melon Fruit
摘要
Abstract
[Objective] In order to make clear the regulatory function of CBF gene in the cold tolerance of Hami melon fruits,the transcription factor named CmCBF1 was cloned and expression analysis was carried out.[Method] Xinmi No.3'Hami melon postharvest fruit was selected as materials.The primers used to amplify the full-length coding sequences were designed based on the predicted CBF1 gene sequence (XM_008440940).These sequences were used as the templates in blast searches against the melon genome databases (https://melonomics.net),and the ATG and TAA codons of the coding regions were determined.Specific primers were designed based on the conserved regions of CBF1,The resulting cDNA was employed as a template for PCR reactions using gene-specific primers to amplify the coding regions of CmCBF1,the expression of CmCBF1 was carried out in triplicate with different cDNA samples synthesized from tissue collected at each sampling time with PCR amplification.[Result]The CmCBF1 gene from the cDNA contained an open reading frame of 639 bp,its GenBank accession number was KT737742,which encoded a polypeptide of 212 amino acids residues with a molecular mass of 23.89 kD and a pI of 5.23.The sequence homology comparison showed that the CmCBF1 had a relatively close evolutionary relationship with Fragariaananassa,FaCmCBF1,which shared 85 ％.RT-PCR analysis showed that the expression of CmCBF1 was induced by low temperature,including 1℃ 、3℃ and 5℃,but not expressed at room temperature at 20℃.[Conclusion]There is a negative correlation between the relative expression of transcription factor CmCBF1 and the incidence of chilling injury of Hami melon fruit,which provides the theoretical basis for studying the mechanism of the incidence of chilling injury of Hami melon fruit during the storage at low temperature.关键词
哈密瓜/CmCBF1/克隆/表达Key words
Hami melon fruit/CmCBF1/gene cloning/gene expression分类
农业科技引用本文复制引用
张婷,潘俨,孟新涛,郑素慧,徐斌..哈密瓜果实转录因子CmCBF1的克隆及表达分析[J].新疆农业科学,2017,54(10):1765-1774,10.基金项目
新疆维吾尔自治区自然科学基金项目“哈密瓜抗冷途径中CBF转录因子基因的克隆与表达”(2014211B026)The Natural Science Foundation of Xinjiang,China "Cloning and expression analysis of a CRT/DRE-binding factor gene CBF from Hami melon fruit during cold resistance pathway" (2014211B026) （2014211B026）
