表达盒两侧不同核基质结合区序列介导表达载体的构建

Construction of the expression vector which contains two different matrix attachment regions flanking the expression cassette

目的构建表达盒两侧含有不同来源核基质结合区(MAR)序列的载体pCCF-βg,为进一步研究MAR的调控功能提供基础。方法用限制酶酶切质粒pCCM-in,将切下的interferon-MAR片段连接至表达盒一侧含有globin-MAR的表达载体上,构建表达盒两侧含异源MAR的表达载体pCCF-βg。结果酶切及琼脂糖凝胶电泳证实所构建的载体pCCF-βg正确,pCCF-βg表达盒两侧分别含有interferon-MAR和globin-MAR片段。结论成功构建了表达盒两侧含有不同来源的MAR片段的载体pCCF-βg。

Objective To construct the expression vector pCCF-βg which contains two different matrix attachment regions （MARs） flanking the expression cassette,in order to study the regulation function of MAR further. Methods The vector pCCM-in was cut by restriction enzymes,interferon-MAR was recovered and ligated into the vector pCCM3-βg which contains globin-MAR, the pCCF-βg vector which contains two different MARs flanking the expression cassette was constructed. Results The pCCF-βg vector which contains globin-MAR and interferon-MAR flanking the expression cassette was proved to be right through digestion with the corresponding enzymes and agarose gel electrophoresis. Conclusion The expression vector pCCF-βg which contains the different MARs flanking the expression cassette was successfully constructed.