果蝇新型碱性磷酸酶相关基因CG6236的敲除及表型分析  

Drosophila Novel Alkaline Phosphatase Related Gene CG6236 Deletion and Phenotypic Analysis

作　　者：缪海锋[1] 方可欣[1] 黄爽爽[2] 金龙金[2] 林鑫华[3] 吴一卉[3] 

机构地区：[1]温州医学院眼视光学院,温州325027 [2]温州医学院检验医学院,生命科学学院,温州325035 [3]中国科学院动物研究所生物膜与膜生物工程国家重点实验室,北京100101

出　　处：《中国细胞生物学学报》2012年第5期424-431,共8页Chinese Journal of Cell Biology

基　　金：国家重大科技计划专项课题(No.2011CB943900);温州医学院人才启动项目(No.QTJ08012);温州医学院科研基金重大项目(No.XNK07005);温州市科技局计划项目(No.Y20110154);温州医学院眼视光医院课题(No.YNKT201113)资助项目~~

摘　　要：碱性磷酸酶广泛存在于人体各器官中,其水平异常与一系列疾病有关,但其在发育中的具体作用机制尚不明确。该文以果蝇为模式生物,研究一个新型碱性磷酸酶样蛋白基因CG6236在发育中的功能。利用P-因子介导的不精确剪切获得CG6236缺失突变体果蝇品系,发现CG6236纯合突变体半致死,存活的幼虫及成蝇腹部出现黑色素瘤样表型。在细胞中表达CG6236融合蛋白,发现其定位于细胞质中。另外,缺失或过表达CG6236均不影响Wingless和Hedgehog信号通路。综上,该研究首次获得CG6236基因缺失突变和转基因果蝇品系,并观察了缺失CG6236果蝇的表型,为进一步阐明基因CG6236的功能及作用机制奠定了基础。Alkaline phosphatase（ALP） is present in all tissues through the entire body in human,and the abnormal ALP level is related to many diseases.However,the molecular mechanism of ALP during development remains unclear.Here,we report and analyze the function of gene CG6236,an important ALPlike gene,using Drosophila model system.We generated 3 mutant alleles for CG6236 using imprecise P-element excision method.We found that homo-mutant alleles were semi-lethal,and there were melanotic tumors in the abdomen of homozygous mutant larvaes and adult flies.Moreover,we generated pUAST-CG6236-V5 construct and its transgenic lines.The staining results showed that the protein of CG6236 was localized in the cytoplasm.Furthermore,by mosaic clone analysis and over-expression of CG6236 in wing discs,we showed that CG6236 is not required for Wingless and Hedgehog signaling.Taken together,this is the first time to generate CG6236 mutants and transgenic lines in Drosophila,and uncover the phenotype of a novel ALP related gene CG6236.Our findings provided evidence and laid foundation for further research to elucidate the physiological function and molecular mechanism of CG6236.

关 键 词：CG6236 碱性磷酸酶 基因缺失 黑色素瘤 果蝇 

分 类 号：R346[医药卫生—基础医学]