Clarification of the role of annexin on glucose transport

最近我们发现了一种新的Akt结合蛋白，Annexin，它的特征是依赖于钙离子与细胞膜的相互作用。首先，利用HEK293细胞的cDNA文库，根据已报道的序列，通过聚合酶链式反应(PCR)扩增出编码人膜联蛋白的cDNA，构建了表达人膜联蛋白的重组腺病毒。我们用腺病毒系统证实了膜联蛋白与Akt的结合，并用该抗原免疫兔制备了GST-Annexin融合蛋白，以产生针对膜联蛋白的抗体。以全长cDNA为模板，通过聚合酶链式反应设计Akt和Annexin基因片段，构建GST融合蛋白。

Recently we have discovered a novel Akt binding protein, annexin, which is characterized by Ca2+-dependent interactions with cellular membranes. First we produced cDNA encoding human annexin by PCR with primers corresponding to sequences already reported, using the cDNA library from HEK293 cells and generated recombinant adenovirus to express human annexin. We confirmed the association of annexin with Akt using adenovirus system and then made GST-annexin fusion protein to generate the antibody specific against annexin by immunizing rabbits with this antigen. Fragments of both Akt and annexin were designed by PCR using full-length cDNAs as templates to generate GST-fused proteins.