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2025年5月10日 星期六
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猪心型脂肪酸结合蛋白基因的克隆及不同生长阶段表达的差异

黄玉海;汪以真;单体中;刘建新;冯杰
浙江大学饲料科学研究所,教育部动物分子营养学重点实验室, 杭州  310029
Cloning of Heart-type Fatty Acid-binding Protein (H-FABP ) Gene of Swine and
Difference of  Its Expression at Different Stages
HUANG Yu-hai;WANG Yi-zhen;SHAN Ti-zhong;LIU Jian-xin;FENG Jie
Institute of Feed Science, Zhejiang University, Key Laboratory of Molecular Animal Nutrition, Ministry of Education, Hangzhou 310029, China
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摘要 从1, 30, 50, 70和90 kg左右的杜长大猪肌肉组织中提取基因组RNA,用RT-PCR扩增猪心型脂肪酸结合蛋白(H-FABP)基因,获得1条211 bp的片段,以pGEM-T  为载体,将该基因片段克隆到大肠杆菌(Escherichia coli ) DH5α中。从筛选到的阳性克隆中分离出脂肪酸结合蛋白基因,测定其序列。分析表明,该片段为脂肪酸结合蛋白基因cDNA的部分序列,与已报道的猪肌肉组织中的H-FABP cDNA部分序列同源性达到99%。以H-FABP基因片段的克隆为基础,构建了优化的半定量RT-PCR法,以18S rRNA为内标,研究不同生长阶段猪肌肉组织中H-FABP基因表达的差异。结果表明,从出生到50 kg,猪H-FABP基因表达呈下降趋势;50~90 kg阶段, H-FABP基因的表达又有所上升。
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黄玉海
汪以真
单体中
刘建新
冯杰
关键词 心型脂肪酸结合蛋白基因 基因克隆基因表达    
AbstractHeart-type fatty acid-binding protein(H-FABP) gene mRNA extracted from muscle of pig(Duroc×Landrace×Yorkshire) was amplified using RT-PCR. And a DNA fragment of 211 bp in length was obtained and the PCR product was cloned into PGEM-T vector. The H-FABP gene was isolated and sequenced from the positive clones screened. Sequence analysis suggested that this fragment was partial sequence of H-FABP cDNA. The gene homology of fragment obtained in this study compared with that of reported H-FABP sequence in muscle of porcine was 99%. Based on the H-FABP gene clone, an optimal semi-quantitative RT-PCR method was successfully constructed. Using 18S  rRNA as inner control, the difference of H-FABP gene expression at different stages  of swine was researched. The result indicated that the H-FABP gene expression level decreased from 1 to 50 kg, and increased from 50  to 90 kg of  body weight.
Key wordsswine    heart-type fatty acid-binding protein    gene clone    gene expression
收稿日期: 2005-10-20     
ZTFLH:  S188  
通讯作者: 汪以真   
引用本文:   
黄玉海;汪以真;单体中;刘建新;冯杰.

猪心型脂肪酸结合蛋白基因的克隆及不同生长阶段表达的差异

[J]. , 2006, 14(5): 657-661.
HUANG Yu-hai;WANG Yi-zhen;SHAN Ti-zhong;LIU Jian-xin;FENG Jie. Cloning of Heart-type Fatty Acid-binding Protein (H-FABP ) Gene of Swine and
Difference of  Its Expression at Different Stages
. , 2006, 14(5): 657-661.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/      或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2006/V14/I5/657
 
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