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huojinlong8610

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【摘要】目的 获得版纳微型猪近交系生长激素受体(Growth hormone receptor,GHR)基因序列,通过生物信息学分析预测GHR功能并进行GHR mRNA多组织表达谱分析。方法 以版纳微型猪近交系的肝脏组织为材料提取RNA,RT-PCR方法扩增GHR基因编码区序列,将序列连接至pMD18-T载体进行克隆、测序和生物信息学分析;半定量方法检测GHR mRNA在BMI不同组织中表达量的差异。结果 成功克隆了BMI GHR 编码区序列,提交GenBank获得登录号KC999114。该基因CDS长1917 bp,编码638个氨基酸。生物信息学分析表明,与长白猪的GHR序列相比BMI存在4处氨基酸替换,分别为p. E381D,p. A409S,p. L556V和p. A580G,均发生在GHBP区。GHR基因多组织表达谱分析显示:GHR mRNA几乎在各组织中均有表达,其中在神经纤维、心、肝、脾、肺、小肠、卵巢和肌肉中表达量都很高,在大脑、肾、胃和胰中的表达量较低。结论 GHR的氨基酸替换可能会影响GHR与GH的结合,导致其体型矮小。
[Abstract] Objective To get the BMI GHR gene sequence, predict its function by bioinformatics analysis and obtain the GHR mRNA tissues transcription profile. Methods BMI GHR cDNA sequence was cloned from liver RNA by RT-PCR. Inserted the product into pMD18-T vector for clone, sequencing and bioinformatics analysis. Conducted semi-quantitative RT-PCR to determine its expression in different tissues. Results The GHR cDNA sequence was cloned and got the GenBank Accession No. KC999114. The encoding sequence was 1917 bp and encoded a protein of 638 amino acids. Bioinformatics analysis showed that there were four amino acids substitutions were found in the GHR binding protein superfamilies between BMI and Landrace pig GHR protein. The substitutions were p. E381D, p. A409S, p. L556V and p. A580G. GHR mRNA tissues expression analysis revealed that GHR mRNA was expressed in almost all tissues. It was most highly expressed in the nerve fiber, heart, liver, spleen, lung, small intestine, ovary and muscle while weakly expressed in the brain, kidney, stomach and pancreas. Conclusions The BMI GHR substitutions maybe weaken the interaction of GHR and GH which can lead to BMI dwarfism.
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biomed_fanyi

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huojinlong8610: 金币+20, 翻译EPI+1, 有帮助 2013-07-29 17:46:31
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lemon_time

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【答案】应助回帖

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huojinlong8610: 金币+80, ★★★★★最佳答案 2013-07-29 17:46:40
[Abstract] Objective To get the sequence of BMI GHR gene, predict its function by bioinformatics analysis and investigate its mRNA expression profile in different tissues. Methods BMI GHR cDNA sequence was cloned from liver RNA by RT-PCR. Then the products were inserted into pMD 18-T vector for cloning, sequencing and bioinformatics analysis. It was to determine GHR mRNA expression in different tissues by conducting semi-quantitative RT-PCR. Results The GHR cDNA sequence was cloned and the the GenBank Accession No. was KC999114. The encoding sequence was 1917 bp and encoded a protein of 638 amino acids. The bioinformatics analysis result showed that there were four amino acids substitutions at the GHBP superfamilies of GHR protein comparing BMI with Landrace. The substitutions were p. E381D, p. A409S, p. L556V and p. A580G. Tissues expression analysis revealed that GHR mRNA was expressed in almost all tissues. It was most highly expressed in the nerve fiber, heart, liver, spleen, lung, small intestine, ovary and muscle while weakly expressed in the brain, kidney, stomach and pancreas. Conclusions The GHR substitutions may affect the interaction of GHR and GH which can infulence the BMI growth and development.
3楼2013-07-29 17:44:51
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