一种新的遗传性凝血因子ⅩⅢ缺乏症ⅩⅢA亚基mRNA大片段缺失的鉴定  

Identification of a novel large deletion of factor ⅩⅢ subunit A mRNA associated with hereditary factor ⅩⅢ deficiency

作　　者：马秋玲[1] 周克元[2] 周鹏[3] 蔡望伟[1] 

机构地区：[1]海南医学院生物化学与分子生物学教研室,海口571101 [2]广东医学院生物化学教研室 [3]中国热带农业科学院热带生物技术研究所

出　　处：《中华血液学杂志》2012年第4期299-302,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金（30060037）;教育部科学技术研究重点项目（03147）

摘　　要：目的分析1个遗传性凝血因子）ⅩⅢ缺乏症家系白细胞表达的凝血因子）ⅩⅢA亚基mRNA。方法采用RT—PCR、半定量RT—PCR、克隆和测序方法分析遗传性凝血因子ⅩⅢ缺乏症家系先证者及其亲属外周血单个核细胞的凝血因子ⅩⅢA亚基mRNA；用SWISS．MODEL分析凝血因子ⅩⅢA亚基基因突变对蛋白质结构和功能的影响。结果①先证者凝血因子ⅩⅢA亚基mRNA存在第11到第279个密码子大片段缺失（DelCD11-279），该缺失为阅读框内缺失，编码1条454个氨基酸截短的凝血因子]ⅩⅢA亚基；先证者的凝血因子）ⅩⅢA亚基mRNA表达量明显减少；②三维结构分析显示DelCD11—279的凝血因子ⅩⅢA亚基mRNA所编码1条464个氨基酸截短的凝血因子ⅩⅢA亚基蛋白缺乏部分活性肽、B一夹层区和部分催化中心区，不能形成正常的催化中心，缺乏转谷氨酰胺酶活性。结论凝血因子）ⅩⅢA亚基mRNADelCD11—279与遗传性凝血因子）ⅩⅢ缺乏相关，该家系患者凝血因子）ⅩⅢA亚基基因表达水平降低是导致凝血因子）ⅩⅢ缺乏的原因之一。Objective To analyze the expressed mRNA of the factor ⅩⅢ subunit A （FⅩⅢA） in monocyte in a hereditary factor xm （FⅩⅢ） deficiency family. Methods The FⅩⅢA mRNA of the proband and the other family members was analyzed by RT-PCR, semi-quantitative RT-PCR, cloning and sequencing. The three dimensional structure of the protein was predicted by SWISS-MODEL and viewed by RASMIOL. Results （1）A large inframe deletion from codons 11 to 279, spanning from exon 2 to 7 of FⅩⅢA （DelCDll-279）, was identified in the proband at mRNA level and a truncated protein is predicted composed of 464 amino acids. Compared with the normal and the other families, the proband showed lower level of FⅩⅢA mRNA in RT-PCR. （2）SWISS-MODEL analysis showed that the truncated protein lacked the B-sandwich and a part of catalytic core, resulting in loss of the normal catalytic domains. Conclusion DelCDll-279 of FⅩⅢA mRNA is associated with hereditary FⅩⅢ deficiency. The reduced expressing level of FⅩⅢA gene is one of the causes resulting in FⅩⅢ deficiency in the patients.

关 键 词：因子ⅩⅢ 因子ⅩⅢ缺乏 DNA突变分析 

分 类 号：R55[医药卫生—血液循环系统疾病]