Cloning and bioinformatics analysis of GHR gene in Banna minipig inbred line
Received:September 25, 2013  
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KeyWord:Banna minipig inbred line (BMI);Growth hormone receptor (GHR);Bioinformatics analysis
王淑燕 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
霍金龙 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
潘伟荣 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
施晨 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明
曾养志 云南农业大学 云南省版纳微型猪近交系重点实验室, 昆明 ;云南农业大学 动物科学技术学院, 昆明
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      Objective To get the Banna minipig inbred line (BMI) GHR gene sequence, predict its function by bioinformatics analysis and obtain the GHR mRNA multi-tissues transcription profile. Methods BMI GHR cDNA sequence was cloned from liver RNA by RT-PCR. Then the products were inserted into pMD18-T vector for cloning, sequencing and bioinformatics analysis. The GHR mRNA expression in different tissues was determined by semi-quantitative RT-PCR. Results The GHR cDNA sequence was cloned and the GenBank accession No. was KC999114. The length of encoding sequence was 1917 bp and encoded a protein of 638 amino acids. The results of bioinformatics analysis showed that four amino acid substitutions were found at the intracellular domain of GHR between BMI and Landrace pigs. The substitutions were p. E381D, p. A409S, p. L556V and p. A580G. Analysis of the GHR mRNA expression in various tissues revealed that it was expressed in almost all tissues. It was most highly expressed in the muscle, moderate in small intestine, heart, liver, nerve fiber, spleen and ovary while weakly expressed in the lung, stomach, cerebrum, pancreas and kidney. Conclusions We have cloned complete GHR coding sequence, performed bioinformatics analysis and tissue expression profile analysis. It provides the primary foundation for further insight into the dwarf mechanism of BMI.
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