猪G蛋白偶联受体12-Gpr12-基因克隆鉴定及其表达分析

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1、·10·AnimalHusbandry＆VeterinaryMedicine2014Vol.46No.12猪G蛋白偶联受体12(Gpr12)基因克隆鉴定及其表达分析1，2222*张宝乐，李晔，石放雄，徐银学(1．徐州医学院神经生物学教研室，江苏徐州221002;2．南京农业大学动物科学技术学院，江苏南京210095)摘要:为研究猪G蛋白偶联受体12(Gprotein-coupledreceptor12，Gpr12)基因的序列和组织表达特征，探索Gpr12mＲNA在卵泡发育过程中的表达规律，本研究利用克隆测序技术获取猪Gpr12基因的编码区序列，并利用生

2、物信息学方法分析Gpr12基因序列和蛋白的潜在特征;ＲT－PCＲ技术检测Gpr12基因的组织表达模式;荧光定量PCＲ技术检测Gpr12mＲNA在猪卵泡发育过程中的表达规律。结果表明:猪Gpr12基因编码区序列长1005bp，编码蛋白含有典型的7次跨膜结构域和DＲY基序，存在多个潜在的磷酸化和糖基化修饰位点;Gpr12基因在所检测的组织和细胞中均有表达，且在脑、垂体和卵母细胞中高表达;随着卵泡的发育，Gpr12mＲNA的表达量极显著升高(P＜0.01)，表明Gpr12基因可能参与了猪卵泡发育的调控过程。关键词:猪;G蛋白偶联受体12;基因克隆;表达中图分

3、类号:S828文献标志码:A文章编号:0529－5130(2014)12－0010－05Molecularcloning，identificationandexpressionofGpr12geneinpigs1，2222*ZHANGBaole，LIYe，SHIFangxiong，XUYinxue(1．DepartmentofNeurobiology，XuzhouMedicalCollege，Xuzhou221002，China;2．CollegeofAnimalScienceandTechnology，NanjingAgriculturalUnive

4、rsity，Nanjing210095，China)Abstract:Tostudythesequencefeature，tissueexpressionprofileandtheexpressionpatternintheprocessoffolliculardevelopmentofporcineGprotein-coupledreceptor12(Gpr12)gene，thecompletecodingsequenceofporcineGpr12genewasclonedbyＲT－PCＲ．Bioin-formaticsmethodswereado

5、ptedtoanalyzethesequencefeatureofGpr12geneandpredictthestructureandfunctionofGpr12protein．TheexpressionpatternsofthegeneinvarioustissuesandtheprocessoffolliculardevelopmentwereinvestigatedusingＲT－PCＲandreal-timePCＲ，respectively．TheresultsindicatedthatthecodingsequenceofGpr3genew

6、ithitslengthbeing1005bpnucleotidesencodeda330ami-noacidprotein，whichcontainedthetypicalseven-transmembranestructureofGPCＲsandDＲYmotif，includingseveralpotentialphospho-rylationandN－glycosylationsites．Gpr12genewasexpressedinallthetestedtissuesandcells，theexpressionlevelsofthisgene

7、inbrain，pituitaryandoocytesbeinghigherthanthatinothertissues．TheexpressionlevelofGpr12mＲNAwasup-regulatedsignificantlyduringfolli-cledevelopment(P＜0.01)，suggestingthatGpr12mightbeinvolvedintheregulatoryprocessofporcinefolliculardevelopment．Keywords:pig;Gprotein-coupledreceptor12

8、(Gpr12);molecularcloning;expression［2－3］G蛋白偶联受体