心脏发育候选基因AHNAKβ的克隆和表达研究

Studies on Expression of Heart Developmental Candidate Gene AHNAKβ

心脏发育过程是一个错综复杂的过程,由一系列的基因参与完成。虽然目前已经鉴定出很多与心脏发育相关的基因,但是仍有很多心脏发育相关基因有待鉴定。作者从小鼠心脏cDNA文库中分离并鉴定了一个心脏发育候选基因AHNAKβ。AHNAKβ位于11q12.2,长1 064 bp,由6个外显子和5个内含子组成,其中开放阅读框(ORF)长450 bp(258～710 nt),编码含有149个氨基酸,蛋白质大小约为16.0 kD。我们构建了原核细胞表达载体,在大肠杆菌中表达并纯化了GST-AHNAKβ融合蛋白,然后制备了该蛋白的兔免疫血清。利用该抗体进行了小鼠成体组织Western blot以分析该基因的蛋白表达模式。研究结果表明AHNAKβ在小鼠成体子宫、小肠等多种组织表达,在心脏中表达较高,提示它可能在心脏组织中具有某种重要作用。

The process of heart development is quite complicated, which is regulated by a series of genes. Although many of the heart developmental genes have been identified, there are still a lot of them to be identified. With the aim of identifying the genes involved in human heart development and cardiovascular diseases, we have isolated a heart developmental candidate gene AHNAKβ from a mouse heart cDNA library. AHNAKβ is mapped to chromosome 11 ql2.2. The full-length cDNA is 1 108 bp long and contains an open reading frame （ORF） of 450 nucleotides, which encodes a deduced 16.0 kD protein consisting of 149 amino acids. In order to study the AHNAKβ gene expression patterns, we generated polyclonal anti-AHNAKβ antibodies by injecting a GST fusion protein. In the present study, we show the expression patterns of AHNAKβ in mouse by Western blot analysis. The results of these experiments demonstrated that AHNAKβ has expressed in the heart. In conclusion, these results suggest that a role for AHNAKβ in the regulation of heart development.