小鼠LPAAT基因的克隆和鉴定

发布时间：2018-03-18 02:21

本文选题：溶血磷脂酸脂酰基转移酶　切入点：表达分布模式　出处：《苏州大学》2007年硕士论文　论文类型：学位论文

【摘要】： 目的:克隆与鉴定一个新的小鼠溶血磷脂酸酰基转移酶基因。方法:利用种间同源序列筛选方法克隆了一个新的小鼠溶血磷脂酸酰基转移酶基因,命名为LPAAT-ζ。通过比较基因组DNA序列和cDNA序列,分析和确定了小鼠LPAAT-ζ的基因组结构。利用生物信息学方法分析了LPAAT-ζ蛋白的性质以及小鼠LPAAT家族成员间的关系。利用竞争性半定量RT-PCR的方法,分析了LPAAT-ζ在小鼠不同组织区域的表达分布。利用绿色荧光标记的方法,分析了LPAAT-ζ蛋白在细胞内的定位情况。结果:基因组定位分析显示,小鼠LPAAT-ζ位于第8号染色体的8p24.6区段。小鼠的LPAAT-ζ基因至少由12个外显子组成,在基因组上的跨度至少为17.584kb。LPAAT-ζ基因在小鼠7种组织中的表达分布情况,在脑和肌肉组织中表达量最高,在睾丸中表达量最低,在其余组织中表达量中等。细胞定位结果显示,LPAAT-ζ蛋白定位于细胞质。结论:这一小鼠LPAAT家族新成员的克隆与鉴定,为更全面地研究LPAAT的生物学功能提供了重要的参考价值。
[Abstract]:Objective: to clone and identify a novel mouse lysophosphatidyl transferase gene. Methods: a novel mouse lysophosphatidyl transferase gene was cloned by interspecific homologous sequence screening. The gene was named LPAAT- 味. The genomic DNA sequence and cDNA sequence were compared. The genomic structure of mouse LPAAT- 味 was analyzed and determined. The properties of LPAAT- 味 protein and the relationship among the members of mouse LPAAT family were analyzed by bioinformatics. The competitive semi-quantitative RT-PCR method was used. The expression and distribution of LPAAT- 味 in different tissues of mice were analyzed, and the localization of LPAAT- 味 protein in the cells was analyzed by green fluorescent labeling. Results: genomic localization analysis showed that LPAAT- 味 was located in 8p24.6 region of chromosome 8. The mouse LPAAT- 味 gene was composed of at least 12 exons, and its span was at least 17.584 kb. LPAAT- 味 gene was expressed in 7 kinds of tissues. The expression of LPAAT- 味 protein was the highest in the brain and muscle, the lowest in the testis and the middle in the other tissues. The results of cell localization showed that the LPAAT- 味 protein was located in the cytoplasm. Conclusion: the cloning and identification of a new member of the mouse LPAAT family provides an important reference value for more comprehensive study of the biological function of LPAAT.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R346

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