中文摘要：

目的：建立钙蛋白酶抑制蛋白（calpastatin ，CAST）基因过表达Balb／C小鼠的培育方法，并探讨碱性裂解液提取基因组DNA在子代鼠基因型鉴定中的价值。方法：将雄性C57BL／6‐CAST＋／－转基因小鼠与雌性Balb／C野生型小鼠杂交，将雄性CAST＋／－杂合子代小鼠与雌性Balb／C野生型子代小鼠杂交，连续杂交至消除C57BL／6小鼠遗传背景。取子代小鼠鼠尾组织，采用碱性裂解液提取基因组 DNA ，PCR法扩增 CAST 基因片段，琼脂糖凝胶电泳鉴定结果。采用柯萨奇B3病毒（CVB3）感染Balb／C‐CAST －／－小鼠以建立急性病毒性心肌炎小鼠模型。结果：C57BL／6‐CAST＋／－转基因小鼠与雌性Balb／C野生型小鼠连续杂交10代可消除C57BL／6小鼠遗传背景。采用碱性裂解液提取的基因组DNA数量、纯度能够满足后续实验需要，且产物大小与目的片段一致。成功建立Balb／C‐CAST －／－基因型小鼠急性病毒性心肌炎模型。结论：成功建立Balb／C‐CAST基因过表达小鼠，为后续研究奠定了基础；碱性裂解液提取基因组DNA简便、高效，值得推广。

英文摘要：

Objective： To establish breeding methods of calpastatin （CAST ） gene overexpression in genetic background of Balb/C mice , and to investigate the application value of alkaline lysis solution extracted genomic DNA in genotyping .Methods：The C57BL/6‐CAST＋ /- male mice were hybridized with Balb/C wild type female mice .In the filial generations ,CAST＋ /- heterozygous male mice were hybridized with Balb/C wild type female mice ,continuous hybridization until C57BL/6 genetic background was removed .The genomic DNA was extracted from mice tails ,and CAST gene fragment was amplified by PCR ,the results were identified by agarose gel electrophoresis .The Balb/C‐CAST -/- mice were infected with coxsakievirus（CVB3）to establish the mouse model of acute viral myocarditis .Results：The C57BL/6 genetic background can be removed by the method of C57BL/6‐CAST＋ /- mice countinously hybridized with Balb/C wild type female mice for 10 generations .The alkaline lysis solution extracted genomic DNA amount ,purity can fulfill the needs of following experiments , and the sizes were accordance with target fragment .The Balb/C‐CAST -/- micemodel of acute viral myocarditis was succesfully established .Conclusions：The Balb/C‐CAST gene overexpression transgenic mice is establishes the foundation for following experiments ,the use of alkaline lysis solution to extract genomic DNA is easy ,effective ,which is worth for promotion .