动物营养学报Issue(2)：485-497,13.DOI:10.3969/j.issn.1006-267x.2016.02.022
半滑舌鳎乙酰辅酶 A 羧化酶α基因全长 cDNA分子克隆及饲料脂肪水平对其在肝脏中表达的影响
Full Length cDNA Molecular Cloning of Acetyl-CoA Carboxylase αGene and Effects of Dietary Lipid Level on Its Expression in the Liver of Half-Smooth Tongue Sole (Cynoglossus semilaevis)
摘要
Abstract
The full-length cDNA of acetyl-coa carboxylase α( ACC1 ) gene was cloned from liver of half-smooth tongue sole ( Cynoglossus semilaevis) by reverse transcription PCR ( RT-PCR) and rapid amplification of cDNA ends ( RACE ) methods.The expression of ACC1 mRNA in gut, liver, muscle, ovary, spleen, brain, kidney, heart, mesenteric adipose tissue of half-smooth tongue sole was analyzed by real-time fluores-cence quantitative PCR ( RT-qPCR) method.In addition, the effects of dietary lipid level on ACC1 gene ex-pression in liver of half-smooth tongue sole were investigated.The results showed as follows:1) the full-length cDNA of ACC1 gene was 7 811bp with a 7 074 bp open reading frame encoding 2 357 amino acids.The ACC1 protein has a calculated molecular weight of 266 ku and isolectric point of 6.42.Some conserved sites of ACC1 amino acid sequence were found, including a ATP-binding site ( Gly316 to Gly321 ) , a biotin-binding site ( Val785-Met786-Lys787-Met788 ) , and a CoA-binding site ( Ser1969 to Val1995 ) .In addition, the alternative splice varieties were found in ACC1 gene, and we present evidence for the presence of two isoforms of ACC1 in half-smooth tongue sole liver that differ from the 266 ku ACC1 by the absence of 8 and 15 amino acids.2) The ex-pression of ACC1 mRNA was detected in all examined tissues.The relative expression level of ACC1 mRNA in liver and brain were 2.67 and 2.53, respectively, which were significantly higher than that in other tissues ( P<0.05);the relative expression level of ACC1 mRNA in gut, mesenteric adipose and ovary were second, which were 1.14, 1.10 and 0.97, respectively; the relative expression level of ACC1 mRNA in kidney was the low-est, only was 0.48.3) Compared with the control group ( without fish oil group) , diet added with 3.5%fish oil significantly decreased the relative expression level of ACC1 mRNA in liver ( P<0.05); diet added with 7.0%and 10.0%fish oil led to further decrease the relative expression level of ACC1 mRNA in liver, and it was significantly lower than that in 3.5%fish oil group and control group ( P<0.05) , meanwhile, the 10.0%fish oil group was lower than 7.0%fish oil group ( P>0.05 ) .In summary, we have cloned the full-length cDNA of ACC1 gene from half-smooth tongue sole.Compared with other vertebrates, the main functional sites ( ATP-binding site, biotin-binding site and CoA-binding site) are basically conserved.The lipogenic tissues, such as liver and brain, are the main ACC1 gene expressing tissues in half-smooth tongue sole.Moreover, liver ACC1 mRNA expression is inhibited after the fish fed diets added with fish oil, and the inhibitory effect is posi-tively correlated with the addition of fish oil.关键词
半滑舌鳎/乙酰辅酶A羧化酶α/分子克隆/营养调控/组织表达Key words
half-smooth tongue sole ( Cynoglossus semilaevis)/ACC1/molecular cloning/nutritional regula-tion/tissue express分类
农业科技引用本文复制引用
张夏青,程汉良,许建和,潘茜,易乐飞,彭永兴,申欣,阎斌伦,高焕,王雯祥..半滑舌鳎乙酰辅酶 A 羧化酶α基因全长 cDNA分子克隆及饲料脂肪水平对其在肝脏中表达的影响[J].动物营养学报,2016,(2):485-497,13.基金项目
国家自然科学基金（31272636）；江苏省高校自然科学研究重大项目（10KJA240002）；江苏省自然科学基金（BK2012664）；江苏省海洋生物技术重点实验室开放基金（2009HS15）；浙江省重大科技专项（2012C12907-2）；江苏省优势学科建设工程项目（PAPD）；国家科技支撑计划 （）
