大鼠脑组织中谷氨酸脱羧酶65基因的克隆和序列测定

Cloning and sequencing of glutamic acid decarboxylase 65 gene from rat brain

目的探讨克隆大鼠脑组织中谷氨酸脱羧酶65基因。方法采用RT-PCR方法,将大鼠脑组织中的mRNA逆转录成cDNA,再以cDNA为模板,扩增谷氨酸脱羧酶65基因片段,克隆入T载体,并经序列测定。结果RT-PCR法扩增出一特异产物与预期长度1758bp相符,T载体克隆测序与大鼠谷氨酸脱羧酶65100%同源。结论采用RT-PCR和T载体技术获得了大鼠脑组织中的谷氨酸脱羧酶GAD65基因克隆,为该基因的体外表达打下基础。

Objective To acquire the glutamic acid decarboxylase 65 (GAD65) genes from rat brain . Methods Total RNA extracted from rat brain cell was transcripted reversibly into cDNA with random primers. The glutamic acid decarboxylase 65 gene fragments were amplified using PCR method and sequenced using the dideoxy chain termination technique. Results The product of rat brain was the 1758 bp which matched the size of purpose. The sequence of GAD65 was completely homogeneous with the GAD65 sequence reported. Conclusion GAD65 gene had been cloned from rat brain using RT PCR and T vector techniques. The success of cloning of glutamic acid decarboxylase 65 gene lays a good basis for construction of a recombinant expressed.