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Establishment of a hMSH2/hMSH6 protein interaction system and functional evaluation of hMSH2 gene missense mutations / 中华医学遗传学杂志
Chinese Journal of Medical Genetics ; (6): 559-564, 2013.
Artigo em Chinês | WPRIM | ID: wpr-237207
ABSTRACT
<p><b>OBJECTIVE</b>To construct a hMSH2/hMSH6 protein interaction system, and to use it for evaluating missense mutations detected in hMSH2 gene.</p><p><b>METHODS</b>Recombinant plasmids pGADT7-hMSH2, pGBKT7-hMSH6 and 7 recombinant pGBKT7 plasmids with different hMSH6 domains were constructed through genetic engineering. Subsequently, site-directed mutagenesis was used to construct 10 mutant pGADT7-hMSH2 plasmids, which were transformed into yeast AH109. The growth of transformants was observed on a histidine-deficient culture.</p><p><b>RESULTS</b>Both hMSH6 MutSII-V and MutSIII-V could interact with hMSH2 in yeast AH109. Yeast two-hybrid transformants pGADT7-hMSH2/pGBKT7-hMSH6 MutSII-V were used to construct a hMSH2/hMSH6 protein interaction system. Compared with wild-type hMSH2, yeast two-hybrid transformants c.505A>G, c.1168C>T, c.1255C>A, c.1261C>A could grow normally, c.1223A>G, c.1886A>G, c.2108C>A and c.2516A>G grew slowly, c.518T>G and c.1664 delA could not grow in a histidine-deficient medium in yeast AH109.</p><p><b>CONCLUSION</b>A hMSH2/hMSH6 protein interaction system has been constructed with yeast two-hybrid system, which has been used for functional evaluation of hMSH2 gene missense mutations. c.518T>G is a pathological mutation. c.1223A>G, c.1886A>G, c.2108C>A, c.2516A>G may in part affect the hMSH2 function. And c.505A>G, c.1168C>T, c.1255C>A, c.1261C>A were innocuous variants.</p>
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Ligação Proteica / Saccharomyces cerevisiae / Dados de Sequência Molecular / Sequência de Bases / Química / Mutação de Sentido Incorreto / Técnicas do Sistema de Duplo-Híbrido / Motivos de Aminoácidos / Proteínas de Ligação a DNA / Proteína 2 Homóloga a MutS Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Medical Genetics Ano de publicação: 2013 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Ligação Proteica / Saccharomyces cerevisiae / Dados de Sequência Molecular / Sequência de Bases / Química / Mutação de Sentido Incorreto / Técnicas do Sistema de Duplo-Híbrido / Motivos de Aminoácidos / Proteínas de Ligação a DNA / Proteína 2 Homóloga a MutS Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Medical Genetics Ano de publicação: 2013 Tipo de documento: Artigo