人睫状神经营养因子基因的克隆及序列分析

目的：构建1例2型糖尿病足坏疽截肢患者睫状神经营养因子（hCNTF）cDNA的克隆，为进一步研究睫状神经营养因子与2型糖尿病及其并发症发病的关系奠定基础。方法：从糖尿病足坏疽截肢患者坐骨神经组织中提取总RNA，经逆转录-聚合酶链式反应（RT-PCR）扩增hCNTFcDNA完全编码区，扩增产物经纯化后克隆至pMD18-T载体，并进行测序及序列分析。结果：成功构建此患者CNTFcDNA的克隆，重组质粒（pMD18-T／CNTF）经测序与Genebank检索的hCNTFcDNA突变序列（Accession NM_S72921）100％符合。结论：构建的突变CNTFcDNA克隆为进一步研究CNTF与2型糖尿病及其并发症在发病上的关系提供线索。

Clone and Sequence Analysis of Ciliary Neurotrophic Factor Gene

Objective: To set out the foundation for further research of the pathogenic relationship between ciliary neurotrophic factor (CNTF) and type 2 diabetes (T2DM) by cloning the cDNA of human CNTF from a diabetic foot wound. Methods: To extract total RNA of human sciatic nerve tissue from a diabetic patients with foot wound and amplify the cDNA of human CNTF by RT-PCR. The amplified product was purified and cloned into plasmid pMD-18T. The cloned nucleotide was sequenced and analyzed. Results: The cloning of cDNA of human CNTF was successful. The sequence shared 100% homology with that of Genebank (Accession NM_S72921). Conclusion: The constructed clone of mutated cDNA of human CNTF showed that there would be a possible relationship between CNTF and diabetic complications.