中文摘要：

Ndc80复合体属于有丝分裂中纺锤体检验点成分,位于动粒的外层,由Ndc80、Nuf2、Spc24和Spc25组成,与染色体的稳定性紧密相关。本文构建了p EGFP-N1-Spc24表达载体,研究Spc24对肝再生细胞增殖的影响。提取大鼠肝脏总RNA进行反转录,PCR扩增获得Spc24基因,连接p GEM-T载体,双酶切p GEM-T克隆载体及p EGFP-N1表达载体,连接转化感受态细胞JM109。阳性克隆命名为p EGFP-N1-Spc24。荧光显微镜下检测Spc24的表达水平,并用液压转基因将质粒转入大鼠肝脏内,采用Real-time PCR方法检测增殖细胞核抗原（proliferating cell nuclear antigen,PCNA）的表达情况,并计算肝系数的变化。PCR扩增、酶切鉴定和测序结果表明,成功了构建p EGFP-N1-Spc24表达载体。荧光显微镜观察说明该载体能稳定表达大鼠Spc24基因。转基因后PCNA的表达量高于对照,并且肝系数也高于对照组,表明Spc24是肝再生相关基因,促进肝再生中细胞增殖。

英文摘要：

Ndc80 complexes belongs to the mitotic spindle checkpoint composition, is located in the outer layer of the kinetochore, composed of Ndc80, Nuf2, spindle pole body component 24 homolog（Spc24） and spindle pole body component 25 homolog（Spc25）, closely related to the stability of the chromosome. In this article, the pEGFP - N1 - Spc24 expression vector were built, and the effects of Spc24 on cell proliferation in liver regeneration was studied. Total RNA of rat liver was extracted and transcribed. Spc24 gene amplifited by PCR, then hnked with pGEM - T carrier, pGEM - T cloning vector and pEGFP - N1 expression vector were enzymed by double enzymed, then connected and transformed to JM109 ceils. Positive clone named pEGFP - NI - Spc24. Spc24 expression levels were detected by fluorescence microscope. Plasmids were transfected into in the rat liver by hydraulic method. Using real - time PCR to detect the expression of proliferating cell nuclear antigen （PCNA）, and calculate the changes of liver coefficient. The plasmid is confirmed to be constructed as expectation by PCR amplification, enzyme digestionand sequence reaction. Fluorescence microscopy shows that the plasmid can stable expression rat Spc24 genes. Both the expression of PCNA and liver coefficient in experimental group were higher than the control. These findings indicate that Spc24 is liver regeneration related genes, can promote cell proliferation in the liver regeneration.