| 猪骨髓抗菌肽 PMAP-37 基因片段的克隆及不同体重的表达差异 |
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| 引用本文： | 王燕,汪以真.猪骨髓抗菌肽 PMAP-37 基因片段的克隆及不同体重的表达差异[J].农业生物技术学报,2007,15(4):589-594. |
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| 作者姓名： | 王燕 汪以真 |
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| 作者单位： | 浙江大学饲料科学研究所,动物分子营养学教育部重点实验室,杭州,310029 |
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| 基金项目： | 国家自然科学基金;教育部跨世纪优秀人才培养计划 |
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| 摘 要： | 从杜长大母猪的股骨骨髓中提取基因组RNA，用RT-PCR 扩增抗菌肽PMAP-37 基因，获得1 条约282 bp 的片段，以pGEM-T Easy vector 为载体，将该基因片断克隆到大肠杆菌(Escherichia coli )DH5α 中。从筛选的阳性克隆中分离出PMAP-37 基因，测定其序列。分析表明，该片段为PMAP-37 cDNA 的部分序列，编码167 个氨基酸组成的多肽。研究得到的基因片段与报道的猪骨髓抗菌肽PMAP-37 cDNA 部分序列同源性达到98%。以PMAP-37 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，研究不同体重猪骨髓抗菌肽PMAP-37 基因表达的差异。结果发现，从刚出生到20 kg，PMAP-37 基因表达呈上升趋势，在20～40 kg，PMAP-37 基因表达呈下降趋势，40～60 kg，PMAP-37 基因表达又呈上升趋势，在60～90 kg，PMAP-37 基因表达呈下降趋势。
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| 关 键 词： | 猪 骨髓 抗菌肽 PMAP-37 RT-PCR |
| 文章编号： | 1006-1304（2007）04-0589-06 |
| 收稿时间： | 2006-11-07 |
| 修稿时间： | 2007-01-19 |
Cloning of Antimicrobial Peptide PMAP-37 Gene of Pig and Its Difference Expression at Different Avoirdupois Stages |
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| WANG Yan,WANG Yi-zhen.Cloning of Antimicrobial Peptide PMAP-37 Gene of Pig and Its Difference Expression at Different Avoirdupois Stages[J].Journal of Agricultural Biotechnology,2007,15(4):589-594. |
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| Authors: | WANG Yan WANG Yi-zhen |
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| Abstract: | Genome RNA was extracted from bone marrow of pig (Duroc×Landrace× Yorkshire) and antimicrobial peptide PMAP-37 mRNA was amplified using RT-PCR. A DNA fragment about 282 bp in length was obtained and the PCR product was cloned into pGEM-T vector. The PMAP-37 gene was isolated and sequenced from the screened positive clones. Result of sequence analysis showed that this fragment was the partial sequence of PMAP-37 cDNA and coded 167 amino acid residues. The gene homology of the obtained fragment compared with that of reported PMAP-37 gene sequence in bone marrow of porcine was up to 98%. Based on the PMAP-37 gene clone, an optimal semi-quantitative RT-PCR method was successfully constructed. Using 18S rRNA as inner control, the difference of PMAP-37 gene expression at different avoirdupois of pig was researched, which increased from new born to 20 kg, decreased from 20 to 40 kg, increased from 40 to 60 kg and decreased again at the stage of 60 to 90 kg. |
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| Keywords: | PMAP-37 RT-PCR |
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