樟叶越桔肌动蛋白Actin7基因克隆与组织表达  

Cloning and tissue expression of Actin7 gene from Vaccinium dunalianum Wight

作　　者：蒲芝雨 杨玉菊 张安勉 赵平[2] 丁勇[1] PU Zhi-yu;YANG Yu-ju;ZHANG An-mian;ZHAO Ping;DING Yong(Yunnan Provincial Key Laboratory of Forest Biotechnology,Southwest Forestry University,Kunming 650224,China;Key Laboratory of State Forestry and Grassland Administration on Highly Efficient Utilization of Forestry Biomass Resources in Southwest China,Southwest Forestry University,Kunming 650224,China)

机构地区：[1]西南林业大学云南省高校林木生物技术重点实验室,昆明650224 [2]西南林业大学西南地区林业生物质资源高效利用国家林业与草原局重点实验室,昆明650224

出　　处：《西南农业学报》2023年第4期787-795,共9页Southwest China Journal of Agricultural Sciences

基　　金：国家自然科学基金项目(31960073);云南省大学生创新创业训练计划项目(202110677012)。

摘　　要：【目的】研究樟叶越桔肌动蛋白Actin7基因的组织表达特征。【方法】在樟叶越桔三代转录组数据库的基础上,应用PCR技术成功克隆VdActin7基因cDNA和DNA序列。进一步利用qRT-PCR技术检测并分析VdActin7基因在樟叶越桔幼嫩叶片、成熟叶片、花芽、花、绿色果实、红色果实、绿色果梗和红色果梗等组织中的表达情况。【结果】获得VdActin7基因DNA序列1657 bp, cDNA序列1387 bp。VdActin7基因含有3个内含子,分别为93 bp的intron1、89 bp的intron2和88 bp的intron3,完整开放阅读框长1134 bp,编码377个氨基酸,理论分子量为41.70 KD,等电点为5.31,无信号肽和跨膜结构,属于稳定亲水性蛋白。系统进化树分析发现,VdActin7蛋白与同为越橘属的兔眼越橘(Vaccinium ashei)Actin7亲缘关系最近。VdActin7基因密码子偏好性分析显示,总GC含量和同义密码子第三位核苷酸含量(GC3s)分别为48.2%和44.1%,有效密码子数(ENC)为49.28%,高频密码子共有31个。VdActin7与枣树(Ziziphus jujuba)ZjActin7基因相似,均对CUU偏好性最强,GUU次之,但VdActin对CUU的偏好性(3.43)明显强于ZjActin7(2.57)。qRT-PCR结果显示,VdActin7基因表达呈现明显的组织特异性,在嫩叶中高表达,在成熟叶和果梗中表达量较低,而在果实中检测不到表达量或不表达,暗示VdActin7基因可能在嫩叶的发育过程中发挥重要作用。【结论】本文首次报道了樟叶越桔Actin7基因序列及序列特征,并发现VdActin7基因具有组织表达特异性,为进一步研究VdActin7基因功能、表达调控及作用机制奠定了基础。【Objective】The present paper aimed to study VdActin7 gene expression pattern in different tissues of Vaccinium dunalianum Wight.【Method】The cDNA and DNA of VdActin7 gene were successfully cloned by PCR technique based on the transcriptome database of 3 generations of V.dunalianum.The expression of VdActin7 gene in young leaves,mature leaves,flower buds,flowers,green fruits,red fruits,green fruit stems and red fruit stems of V.dunalianum was further detected and analyzed using qRT-PCR technique.【Result】The full length of VdActin7 gene and cDNA were 1657 and 1387 bp,respectively,which contained 3 introns,namely,intron1 of 93 bp,intron2 of 89 bp and intron3 of 88 bp.It had an open reading frame length of 1134 bp,encoding 377 amino acids.The protein VdActin7 with a relative molecular weight of 41.70 KD and a theoretical isoelectric point of 5.31,was a stable hydrophilic protein,without signal peptide and transmembrane domain.Phylogenetic analysis revealed that the VdActin7 was closest to V.ashei Actin7,which also belonged to the Vaccinium family.The codon usage bias analysis of the VdActin7 gene showed that GC contents of codon(GC)and 3rd nucleotide in codon GC3s were 48.2%and 44.1%,respectively,the effective number of codon(ENC)was 49.28%,a total of high-frequency codons was 31.The codon bias was similar to that of Ziziphus,both with the strongest bias for CUU,followed by GUU,and the bias for CUU was significantly stronger in V.dunalianum(3.43)than that in Ziziphus(2.57).qRT-PCR analysis showed that the VdActin7 gene expression showed obvious tissue specificity,with the highest in young leaves while lower expression in mature leaves and flowers,meanwhile no detectable expression or no expression in fruits,suggesting that the VdActin7 gene might play an important role in the development of young leaves.【Conclusion】In the paper,we reported the Actin7 gene sequence and its characteristics in V.dunalianum for the first time,and found that the VdActin7 gene had tissue expression specificity.This laid the

关 键 词：肌动蛋白7 基因克隆 密码子偏好性 组织表达 樟叶越桔 

分 类 号：S663[农业科学—果树学]