家蝇漆酶2基因的克隆及序列特性与表达分析  

Cloning,Sequence Characteristics and Expression Analysis of Lac-2 Gene in Musca domestica

作　　者：吴书东 黄兰英 彭建 国果 修江帆 吴建伟 尚小丽 WU Shudong;HUANG Lanying;PENG Jian;GUO Guo;XIU Jiangfan;WU Jianwei;SHANG Xiaoli(Guizhou Key and Characteristic Laboratory of Infectious Immune and Antibody Engineering,Guiyang,Guizhou 550025;The Key and Characteristic Laboratory of Modern Pathogen Biology,Basic Medical College,Guizhou Medical University,Guiyang,Guizhou 550025;Xiangya Hospital,Central South University,Changsha,Hunan 410008,China)

机构地区：[1]贵州省感染免疫与抗体工程特色重点实验室,贵州贵阳550025 [2]贵州医科大学基础医学院现代病原生物学特色重点实验室,贵州贵阳550025 [3]中南大学湘雅医院,湖南长沙410008

出　　处：《贵州农业科学》2022年第12期98-105,共8页Guizhou Agricultural Sciences

基　　金：贵州医科大学学术新苗项目(19NSP062);贵州省科技计划项目[黔科合基础-ZK(2022)一般377];国家自然科学基金项目(82002180);贵州省高等学校特色重点实验室[黔教合KY字(2020)012]。

摘　　要：【目的】探明家蝇漆酶2(Lac-2)基因序列特性及表达,为深入研究家蝇漆酶的生物学功能奠定基础。【方法】通过PCR扩增家蝇Lac-2基因的全长cDNA序列;利用生物信息学手段分析其核苷酸和蛋白质序列特征;以GAPDH为内参基因,采用实时荧光定量PCR(Real-Time PCR)技术检测Lac-2基因在家蝇不同发育时期及家蝇3龄幼虫脂肪体、唾液腺、前肠、中肠及后肠中的表达特征。【结果】成功克隆家蝇Lac-2基因的cDNA序列,全长为885 bp,编码269个氨基酸,成熟肽编码蛋白的理论分子量为25 kDa,等电点为7.19。时空表达模式分析显示,Lac-2基因在家蝇3龄幼虫中的表达量最高,在家蝇3龄幼虫的各主要组织中以脂肪体为参照,Lac-2基因在中肠表达水平最高,其次是唾液腺及后肠。【结论】克隆得到家蝇木质素酶Lac-2基因,其功能可能与家蝇的生长发育及食物消化相关。【Objective】The sequence characteristics and expression of Musca domestica Lac-2 gene are identified to lay the experimental foundation for in-depth study of the biological function of endogenous laccase in M.domestica.【Method】The expression characteristics of Lac-2 gene in fat body,salivary glands,foregut,midgut and hindgut of the 3^(rd) instar larvae of M.domestica at different developmental stages is detected by means of amplifying full-length cDNA sequence of M.domestica Lac-2 gene by PCR,analyzing its nucleotide and protein sequence feature by bioinformatics technology and using Real-Time PCR by taking GAPDH as a reference gene.【Result】The cDNA sequence of M.domestica Lac-2 gene is successfully cloned and its full-length and coding are 885 bp and 269 amino acids.The theoretical molecular weight and isoelectric point of the mature peptide encoding protein are 25 KDa and 7.19.The expression of Lac-2 gene reaches the highest in the 3^(rd) instar larvae of M.domestica.The expression of Lac-2 gene is up to the maximum in midgut of the 3^(rd) instar larvae of M.domestica,followed bysalivary glands and hindgut in different main tissues of the 3^(rd) instar larvae of M.domestica by taking fat body as a reference.【Conclusion】The function of the Lac-2 genecloned from M.domestica may be related to growing development and food digestion of M.domestica.

关 键 词：家蝇 木质素酶 漆酶 克隆 序列特性 表达分析 

分 类 号：S476.3[农业科学—农业昆虫与害虫防治]