成年小鼠Myomaker基因克隆及组织表达分析

为研究Myomaker基因在成年小鼠骨骼肌中的表达情况，试验从随机抽取的3只健康成年小鼠骨骼肌中提取总RNA，通过PCR方法扩增Myomaker基因的编码区序列（CDS），并对其进行生物信息学分析。此外，利用免疫组织化学和Western blotting法分析Myomaker蛋白在小鼠骨骼肌组织中的定位与表达。结果显示，Myomaker基因CDS区全长666 bp，编码221个氨基酸，同源性比对结果显示，小鼠与斑马鱼、鸡、猫、牛、犬、人、鹦鹉、猪Myomaker基因CDS区同源性分别为71.7%、80.7%、83.7%、83.2%、84.1%、86.4%、80.6%和84.0%，氨基酸同源性分别为75.9%、79.3%、85.5%、85.5%、88.3%、86.9%、81.4%和93.2%。Myomaker基因编码蛋白分子式为C₁₁₅₄H₁₇₆₉N₂₇₅O₂₉₅S₁₈，分子质量为24 ku，等电点为9.14，不稳定性指数为39.22，平均疏水性为0.501。免疫组织化学和Western blotting分析表明，小鼠Myomaker蛋白在健康成年小鼠骨骼肌中存在表达，且表达于部分肌细胞膜上。本试验结果可为Myomaker基因的深入研究奠定基础。

Cloning and Tissue Expression Analysis of Myomaker Gene in Adult Mice

In order to analyze the expression of Myomaker gene in skeletal muscle of adult mice,the total RNA was extracted from the skeletal muscle of three randomly selected healthy adult mice,and the coding sequence (CDS) of Myomaker gene was amplified by PCR method and analyzed by bioinformatics software.In addition,immunohistochemical and Western blotting methods were used to determine the localization and expression of Myomaker protein in skeletal muscle.The results showed that the CDS region of Myomaker gene was 666 bp,encoding 221 amino acids.The sequence homology of the nucleotide of Myomaker gene in mice were 71.1%,80.7%,83.7%,83.2%,84.1%,86.4%,80.6% and 84.0% comparing with Danio rerio,Gallus gallus,Felis catus,Bos taurus,Canis lupus familiaris,Homo sapiens,Nestor notabilis,Sus scrofa,respectively,and the sequence homology of the amino acids were 75.9%,79.3%,85.5%,85.5%,88.3%,86.9%,81.4% and 93.2%,respectively.The formula of Myomaker protein was C₁₁₅₄H₁₇₆₉N₂₇₅O₂₉₅S₁₈,and the molecular weight,theory isoelectric point,instability index and grand average of hydrophobicity was 24 ku,9.14,39.22 and 0.501,respectively.Immunohistochemical and Western blotting analysis showed that the Myomaker protein expressed in skeletal muscle of healthy adult mice,but it just existed in partial myocyte membrane.This study laid a foundation for the further study of Myomaker gene.