Objective: To detect the expression and the genomic sequence of Doc-1R gene in mice. Methods: The gene specific primers were designed and synthesized according to the cDNA sequence of Doc-1R gene. The sequence of Doc-...Objective: To detect the expression and the genomic sequence of Doc-1R gene in mice. Methods: The gene specific primers were designed and synthesized according to the cDNA sequence of Doc-1R gene. The sequence of Doc-1R gene was cloned by nested PCR. The expression of Doc-1R gene was examined by RT-PCR in thirteen kinds of tissues of mice. Results: The mouse Doc-1R gene has been obtained by two times genomic walking, which spans 2787 bp and contains four exons and three introns. All of the splice donor/acceptor site sequences are in accordance with the consensus 揋T-AG?rule. There was expression of Doc-1R gene in the thirteen tissues. Conclusion: The mouse Doc-1R gene was cloned successfully. The expression pattern suggests that Doc-1R gene is a housekeeping gene, which is important to keep the function of tissues and organs.展开更多
目的构建DOC-1R（deleted in oral cancer-1related）的原核表达载体并且纯化其重组蛋白，用于进-步研究DOC-1R蛋白的结构与功能。方法采用基因重组技术构建原核表达载体pGEX—DOC-1R，经DNA测序鉴定后，转化E．coliB[21，IFFG诱导表达...目的构建DOC-1R（deleted in oral cancer-1related）的原核表达载体并且纯化其重组蛋白，用于进-步研究DOC-1R蛋白的结构与功能。方法采用基因重组技术构建原核表达载体pGEX—DOC-1R，经DNA测序鉴定后，转化E．coliB[21，IFFG诱导表达并纯化融合蛋白。结果以菌落PCR及限制性内切酶酶切鉴定得到候选阳性克隆，测序结果表明所得到的克隆序列及开放阅读框架完全正确，IPTG诱导后SDS—PAGE电泳分析表明在40kD处出现特征蛋白表达带，经磁珠亲合层析后Western印迹检测证实得到较高纯度的GST—p14 DOC-1R融合蛋白。结论成功构建了pGEX—DOC-1R的原核表达载体，表达并纯化出GST—p14 DOC-1R融合蛋白，为DOC，1R抗体制备及研究DOC-1R蛋白结合蛋白及其在细胞周期调控中的生物学功能奠定了基础。展开更多
目的对候选抑癌基因DOC-1R((Deleted in oral cancer-1 related,CDK2AP2)相关基因DCT1(DOC-1Rterminal1)进行染色体定位分析并观察其细胞内定位,检测该基因在人类组织中的表达。方法通过辐射杂交细胞系对其进行染色体定位分析,同时构建E...目的对候选抑癌基因DOC-1R((Deleted in oral cancer-1 related,CDK2AP2)相关基因DCT1(DOC-1Rterminal1)进行染色体定位分析并观察其细胞内定位,检测该基因在人类组织中的表达。方法通过辐射杂交细胞系对其进行染色体定位分析,同时构建EGFP-DCT1质粒并转染HeLa细胞,荧光显微镜观察DCT1蛋白在细胞内的定位。此外,应用荧光定量PCR的方法检测该基因在16个正常人类组织中的表达。结果DCT1基因定位于5q31,其编码产物在HeLa细胞中定位于核膜,该基因在正常人类的16个组织中均有表达,其中在脾中表达水平较高。结论DCT1可能为细胞周期调控相关基因,在人类组织中普遍表达。展开更多
文摘Objective: To detect the expression and the genomic sequence of Doc-1R gene in mice. Methods: The gene specific primers were designed and synthesized according to the cDNA sequence of Doc-1R gene. The sequence of Doc-1R gene was cloned by nested PCR. The expression of Doc-1R gene was examined by RT-PCR in thirteen kinds of tissues of mice. Results: The mouse Doc-1R gene has been obtained by two times genomic walking, which spans 2787 bp and contains four exons and three introns. All of the splice donor/acceptor site sequences are in accordance with the consensus 揋T-AG?rule. There was expression of Doc-1R gene in the thirteen tissues. Conclusion: The mouse Doc-1R gene was cloned successfully. The expression pattern suggests that Doc-1R gene is a housekeeping gene, which is important to keep the function of tissues and organs.
文摘目的对候选抑癌基因DOC-1R((Deleted in oral cancer-1 related,CDK2AP2)相关基因DCT1(DOC-1Rterminal1)进行染色体定位分析并观察其细胞内定位,检测该基因在人类组织中的表达。方法通过辐射杂交细胞系对其进行染色体定位分析,同时构建EGFP-DCT1质粒并转染HeLa细胞,荧光显微镜观察DCT1蛋白在细胞内的定位。此外,应用荧光定量PCR的方法检测该基因在16个正常人类组织中的表达。结果DCT1基因定位于5q31,其编码产物在HeLa细胞中定位于核膜,该基因在正常人类的16个组织中均有表达,其中在脾中表达水平较高。结论DCT1可能为细胞周期调控相关基因,在人类组织中普遍表达。
