应用竞争PCR检测丙酸盐对体外培养牛肝细胞丙酮酸羧化酶mRNA水平的影响研究.pdfVIP

第6届家畜内科学学术研讨会论文集 145 应用竞争PCR检测丙酸盐对体外培养牛肝细胞 丙酮酸羧化酶mRNA水平的影响 徐闯．夏成．刘国文，王哲+，李家奎．张乃生，王兴龙．欧阳红生，张永亮 (吉林大学农学部畜牧兽医学院．吉林长春130062) 擅耍：应用PC(丙酮酸羟化酶)基因组与PCcDNA相比包含内含子序列及在其中插入外源DNA序列的方法，改变 胞PCmILNA水平的影响。使单层肝细胞培养液中丙酸钠浓度分别为0mmol／L、1．5mmol／L、2．5mmol／L、3jmmol／L、4．5 板带。结果表明：随着丙酸钠浓度的升高PCmENA水平呈上升趋势。指示肝细胞内PCmRNA的表达水平受培养液中丙 酸钠浓度的影响。 关健词：丙酮酸羧化酶mRNA；竞争PcR；肝细胞；内含子 E仟ectionof onthe of mRNAinvitro PropionateExpressionPyruvateCarboxylase culturebovine Determinated Reverse Hepatocyte byCompetiveTranscription ChainReaction Polymerase )(U Guo-wen，WANG盈e·。LIJia-kui，殂ANG Chuang，XIACheng，UU Nai-sheng， WANG YANG Xing-long，OUHong-sheng，ZHANGYong·liang and 130062, (AgricultureDepartmentofJiLinUniversity,PrologueVeterinaryInstitute,Ji“n．Changchun China) DNA of wasconstructedthe Abstract：Competive through templatepymvatecarboxylase(PC)eDNA methodsofinlxon andinsertion effectof OIlthe of sequence sequence．Then,thepropionateexpression PCmRNAinvitro bovine dcterminatcd culture Was hepatoeytequantitatively bycompetivePCR．Propionate wasaddedintotheculturemediaasvariousconcentrationof0mM．1．5mM，2．5mM。3．5mM，4．5mM，8．5 mMand11．5 for thentotalRNAwasextracted，l℃verseWas Mm，cultured24h，and transcriptiondone,using thesanle to and levelin lⅡimeramplifyobjectivegene e