| 广西巴马小型猪GHRH成熟肽真核表达质粒对小鼠生长效应的影响 |
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| 引用本文： | 李芳芳,;郭亚芬,;兰干球,;潘堂峰.广西巴马小型猪GHRH成熟肽真核表达质粒对小鼠生长效应的影响[J].广西农业科学,2014(12):2264-2269. |
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| 作者姓名： | 李芳芳 ;郭亚芬 ;兰干球 ;潘堂峰 |
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| 作者单位： | [1]广西畜禽品种改良站,南宁530001; [2]广西大学动物科学技术学院,南宁530005; [3]南宁高新技术产业开发区动物卫生监督所,南宁530005 |
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| 基金项目： | 国家自然科学基金项目（30960058） |
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| 摘 要： | 【目的】探讨广西巴马小型猪促生长激素释放激素（GHRH）成熟肽真核表达质粒对小鼠生长效应的影响,为进一步研究广西巴马小型猪GHRH成熟肽的促生长作用及GHRH基因疫苗的高效制备与安全使用提供参考依据。【方法】利用基因重组技术将广西巴马小型猪GHRH成熟肽序列与p EGFP-N1质粒连接构建重组真核表达质粒,然后按0.5μg/g的剂量肌肉注射经25%蔗糖溶液预处理的小鼠,观察记录小鼠体重变化情况,并分别提取重组质粒注射组小鼠的心脏、肝脏、肾脏及注射部位肌肉DNA,再以PCR检测是否存在基因质粒残留。【结果】以构建的p EGFP-GHRH重组质粒注射小鼠后,其外源GHRH成熟肽DNA可在肌肉中获得表达,但并未整合入小鼠基因组中。虽然重组质粒注射组小鼠的末重与空质粒注射组小鼠的差异不显著（P〉0.05）,但在注射后的10-25 d,重组质粒注射组小鼠体重均显著高于空质粒注射组（P〈0.05）。在相对日增重率方面,注射后的前15 d内,重组质粒注射组小鼠的相对日增重率均高于空质粒注射组。【结论】构建的广西巴马小型猪p EGFP-GHRH重组质粒能在一定时间内促进小鼠生长,即GHRH种属差异性小,在动物中有较高的保守性。
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| 关 键 词： | 广西巴马小型猪 促生长激素释放激素(GHRH)成熟肽序列 小鼠 真核表达质粒 生长效应 |
Eukaryotic expression plasmid of GHRH mature peptide from Guangxi Bama Mini-pig and its influence on mouse growth effect |
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| Institution: | LI Fang-fang, GUO Ya-fen, LAN Gan-qiu, PAN Tang-feng (1Guangxi Work Station of Livestock ＆ Poultry Breed Improvement, Nanning 530001, China; 2College of Animal Science and Technology, Guangxi University, Nanning 530005, China; 3Animal Health Supervision Institute of New ＆ High-tech Industrial Development Zone, Nanning 530005, China) |
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| Abstract: | 【Objective】The present experiment was conducted to investigate the influence of mature peptide eukary-otic expression plasmid of growth hormone releasing hormone（GHRH） in Guangxi Bama Mini-pig on mouse growth effect, in order to provide reference for researching the promoting growth effect of GHRH mature peptide on Guangxi Bama Mini-pig, and efficient preparation and safety use of GHRH gene vaccine. 【Method 】By using gene recombination technique, the GHRH mature peptide sequence of Guangxi Bama Mini-pig was linked into plasmid p EGFP-N1 to construct recombinant eukaryotic plasmid p EGFP-GHRH. The mouse pretreated with 25% sugar solution was treated with intramuscular injection with dose of 0.5 μg/g plasmid to record their weight. Meanwhile aiming at mice in injection group of recombinant plasmid, the DNA of their heart, hepar, kidney and muscle in injection site was extracted to detect residue of gene plasmid by using PCR technique. 【 Result 】 The results showed that, when recombinant plasmid p EGFP-GHRH was injected into mouse, the ex-GHRH mature peptide sequence was expressed in the muscle, while it was not integrated into mouse genome. Compared injection group of recombinant plasmid with injection group of empty plasmid, there was no significant difference in final weight of mice between the former and the latter（P0.05）. While during 10-25 days after injection, the weight of the former was significantly higher than that of the latter（P 0.05）.As for relative daily gain rate, during 15 days after injection, the weight of the former was higher than that of the latter.【Conclusion 】The recombinant plasmid p EGFP-GHRH of Guangxi Bama Mini-pig can promote growth of mouse for a certain time. Therefore, GHRH has small difference in different species and high conservation in animal. |
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| Keywords: | Guangxi Bama Mini-pig mature peptide sequence of growth hormone releasing hormone（GHRH） mouse eukaryotic expression plasmid growth effect |
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