小鼠RPL9基因克隆及组织表达差异性研究
|
摘要
根据已报道的小鼠核糖体蛋白L9基因(RPL9)序列设计PCR引物,克隆RPL9基因,并以β-actin为内参基因,采用相对荧光定量RT-PCR方法,检测、分析RPL9基因mRNA在小鼠心、肝、脾、肺、肾、脑及脂肪7个组织中表达量的差异。结果表明,成功克隆出RPL9基因序列全长,该基因在小鼠的7个组织中均有不同程度的转录表达,其中,在脂肪中的表达量最高,在肾、脾和脑中的表达量较高,在心、肝中的表达量较低,在肺中表达量最低。
We designed PCR primers which were based on ribosomal protein L9 gene( RPL9) sequence in mouse reported,and then cloned RPL9 gene. Using β-actin as internal gene,the method of the relative fluorescence quantitative RT-PCR was used to detect and analyze the expression differences of RPL9 gene mRNA in seven kinds of tissues in mouse,such as heart,liver,spleen,lung,kidney,brain and fat. The results showed that we cloned RPL9 gene successfully,the expression of RPL9 gene had different levels of transcription in different tissues,the highest amount of expression in the fat,higher expression in kidney,spleen and brain,lower amount of expression in the heart and liver,the lowest amount of expression in the lung.
We designed PCR primers which were based on ribosomal protein L9 gene( RPL9) sequence in mouse reported,and then cloned RPL9 gene. Using β-actin as internal gene,the method of the relative fluorescence quantitative RT-PCR was used to detect and analyze the expression differences of RPL9 gene mRNA in seven kinds of tissues in mouse,such as heart,liver,spleen,lung,kidney,brain and fat. The results showed that we cloned RPL9 gene successfully,the expression of RPL9 gene had different levels of transcription in different tissues,the highest amount of expression in the fat,higher expression in kidney,spleen and brain,lower amount of expression in the heart and liver,the lowest amount of expression in the lung.
引文
[1]Li D,Chen N,Mc Michael A J,et al.Generation and characterisation of CD1d tetramer produced by a lentiviral expression system[J].Journal of Immunological Methods,2008,330(1):57-63.
[2]Kroes R A,Jastrow A,Mc Lone M G,et al.The identication of novel therapeutic targets for the treatment of malignant brain tumors[J].Cancer Lett,2000,156(2):191-198.
[3]Sahin F,Qiu W,Wilentz R E,et al.RPL38,FOSLl,and UPPl are predominantly expressed in the pancreatic ductal epithelium[J].Pancreas,2005,30(2):158-167.
[4]王林云.中国养猪业:如何迎接WTO的挑战,21世纪养猪业与人类健康[C]//21世纪养猪业与人类健康.北京:中国畜牧兽医学会猪学分会,2001:20-24.
[5]Shuda M,Kondoh N,Tanaka K,et al.Enhanced expression of translation factor mRNAs in hepatocellular carcinoma[J].Anticancer Res,2000,20(4):2489-2494.
[6]许琰,丛喆,魏强,等.实时定量PCR的研究进展及应用[J].中国试验动物学报,2007,15(2):155-158.
[7]黄河,帅素容.藏猪心脏脂肪酸结合蛋白基因(HFABP)组织表达差异性研究[J].四川农业大学学报,2007,25(4):480-484.
[8]齐小辉,马玺,李珊珊,等.基于PCR的基因差异表达分析技术[J].生物技术通讯志,2004,15(4):389-391.
[9]史建伍,盛军庆,曾柳根,等.萍乡红鲫卵母细胞透明带糖蛋白ZP3基因及其侧翼序列的克隆和分析[C].中国南方十六省(市、区)水产学会渔业学术论坛第二十六次学术交流大会论文集(下册),2010.
[10]杨华,刘守仁,钟发刚,等.BMPR-IB基因在绵羊不同组织的表达差异性研究[J].中国畜牧杂志,2009,45(11):6-10.
[11]陈修栋,桂宏翔,赵枭阳,等.DDK综合症部分相关基因的实时定量PCR分析[J].河南农业科学,2013,42(7):141-144,149.
[12]郑梦月,姜冬梅,康波,等.四川白鹅ENO1基因特征及其在HPG轴组织中发育性表达的研究[J].华北农学报,2014,29(1):93-97.
[13]秦彤,王皓宇,郝海生,等.日粮精粗比对围产期奶牛乳腺内参基因表达的影响[J].华北农学报,2013,28(4):75-78.
[14]张萍,周慧芳,孙正华,等.恒河猴CD1d基因的克隆及其组织表达差异性分析[J].细胞与分子免疫学杂志,2009,25(7):581-584.
[15]张晶,陈修栋,王善强,等.淮南猪品种遗传特性的AFLP分析[J].中国畜牧杂志,2011,38(5):130-133.
[16]Gudipati R K,Neil H,Feuerbach F,et al.The yeast RPL9B gene is regulated by modulation between two modes of transcription termination[J].EMBO,2012,31(10):2427-2437.
[2]Kroes R A,Jastrow A,Mc Lone M G,et al.The identication of novel therapeutic targets for the treatment of malignant brain tumors[J].Cancer Lett,2000,156(2):191-198.
[3]Sahin F,Qiu W,Wilentz R E,et al.RPL38,FOSLl,and UPPl are predominantly expressed in the pancreatic ductal epithelium[J].Pancreas,2005,30(2):158-167.
[4]王林云.中国养猪业:如何迎接WTO的挑战,21世纪养猪业与人类健康[C]//21世纪养猪业与人类健康.北京:中国畜牧兽医学会猪学分会,2001:20-24.
[5]Shuda M,Kondoh N,Tanaka K,et al.Enhanced expression of translation factor mRNAs in hepatocellular carcinoma[J].Anticancer Res,2000,20(4):2489-2494.
[6]许琰,丛喆,魏强,等.实时定量PCR的研究进展及应用[J].中国试验动物学报,2007,15(2):155-158.
[7]黄河,帅素容.藏猪心脏脂肪酸结合蛋白基因(HFABP)组织表达差异性研究[J].四川农业大学学报,2007,25(4):480-484.
[8]齐小辉,马玺,李珊珊,等.基于PCR的基因差异表达分析技术[J].生物技术通讯志,2004,15(4):389-391.
[9]史建伍,盛军庆,曾柳根,等.萍乡红鲫卵母细胞透明带糖蛋白ZP3基因及其侧翼序列的克隆和分析[C].中国南方十六省(市、区)水产学会渔业学术论坛第二十六次学术交流大会论文集(下册),2010.
[10]杨华,刘守仁,钟发刚,等.BMPR-IB基因在绵羊不同组织的表达差异性研究[J].中国畜牧杂志,2009,45(11):6-10.
[11]陈修栋,桂宏翔,赵枭阳,等.DDK综合症部分相关基因的实时定量PCR分析[J].河南农业科学,2013,42(7):141-144,149.
[12]郑梦月,姜冬梅,康波,等.四川白鹅ENO1基因特征及其在HPG轴组织中发育性表达的研究[J].华北农学报,2014,29(1):93-97.
[13]秦彤,王皓宇,郝海生,等.日粮精粗比对围产期奶牛乳腺内参基因表达的影响[J].华北农学报,2013,28(4):75-78.
[14]张萍,周慧芳,孙正华,等.恒河猴CD1d基因的克隆及其组织表达差异性分析[J].细胞与分子免疫学杂志,2009,25(7):581-584.
[15]张晶,陈修栋,王善强,等.淮南猪品种遗传特性的AFLP分析[J].中国畜牧杂志,2011,38(5):130-133.
[16]Gudipati R K,Neil H,Feuerbach F,et al.The yeast RPL9B gene is regulated by modulation between two modes of transcription termination[J].EMBO,2012,31(10):2427-2437.