一个新的人乙酰化酶MAK3的克隆、表达和纯化

Cloning, Expression and Purification of a Novel Human GCN5-related Acetyltransferase, MAK3

在对乙酰化酶GNAT家族进行研究分析的过程中,找到一个人GNAT家族的新成员MAK3。MAK3含有一个保守的乙酰化酶结构域,并且不同物种中的MAK3的蛋白质序列高度保守。通过半定量和定量RT-PCR检测了在不同组织中MAK3的mRNA水平。并且把MAK3分别克隆到了真核和原核表达质粒中,通过免疫印迹技术证实了MAK3的表达。还进一步用亲和层析的方法成功纯化了细菌中表达的MAK3,并对纯化后的MAK3的酶活性进行了测定。

A novel human GCN5-related acetyltransferase, MAK3 was found by bioinformatical analysis of the GNAT family. Human MAK3 contains a conserved acetyltransferase domain and the protein sequence of MAK3 is highly conserved in different species. We detected mRNA level of MAK3 in different tissues by semi-quantity and full quantity RT-PCR and cloned MAK3 into both mammalian and bacterial expression vectors, and the expression of MAK3 was confirmed by Western blot. MAK3 expressed in bacteria was further purified with affinity chromatography. Moreover, the enzyme activity of purified MAK3 protein was also detected.