小鼠ZnT3 cDNA片断的克隆及其mRNA表达

Cloning of ZnT3 cDNA and expression of ZnT3 mRNA in mice

为了解ZnT3 (zinctransporter3 )mRNA在小鼠各组织中的表达状况 ,用反转录多聚酶链反应法 (RT PCR)克隆ZnT3cDNA片断 ,荧光标记的全自动测序法确定ZnT3cDNA片断的碱基顺序 ,RT PCR法检测小鼠各组织ZnT3mRNA表达并比较在组织中的表达量。结果 :RT PCR获得单一条带的片断 ,其大小 70 0bp ,碱基顺序与文献报道序列一致 ;在大脑皮层、海马和睾丸中检测到ZnT3mRNA表达 ,其中睾丸中表达量最高 ,大脑皮层、海马表达量无显著性差别 ,心、肝、脾、肺、肾、小肠及嗅球、小脑等组织中未见ZnT3mRNA表达。说明克隆到正确的ZnT3cDNA片断。ZnT3mRNA主要表达于睾丸、大脑皮层、海马 。

In order to determine the level of mouse zinc transporter3 (ZnT3) mRNA expression, the fragment is cloned by RT PCR method and is sequenced by dideoxynucleotide method. The expression of ZnT3 mRNA is also examined by RT PCR method and is compared with β actin that served as the control. As the result, a proper single fragment of 700bp is obtained with the sequence conformable to the corresponding fragment sequence of reported mouse ZnT3 cDNA. The ZnT3 mRNA is detected in cerebrum, hippocampus and in testis in that the highest expression is observed. However, the expression of ZnT3 mRNA is not detected in heart, liver, lung, spleen, kidney, intestine, olfactory bulb and cerebellum. The expression of ZnT3 mRNA in brain and testis suggests a physiologically functional importance of ZnT3 in mental and reproductive activities of mouse.