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猪精氨酸-丝氨酸蛋白激酶1(SRPK1)克隆表达及功能初步分析 被引量:3
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作者 俄广鑫 刘娣 +6 位作者 张冬杰 杨秀琴 崔羽 祝继原 杨少成 王嘉博 谢宇彤 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第11期1379-1386,共8页
本研究旨在阐明猪SRPK1基因的分子特点和表达谱。以大白猪为研究对象,利用RT-PCR技术克隆SRPK1基因CDS区域,同时利用反向PCR(inverse PCR,I-PCR)技术得到猪部分SRPK1的5′UTR序列;利用生物信息学方法分析SRPK1基因核苷酸序列及编码蛋白... 本研究旨在阐明猪SRPK1基因的分子特点和表达谱。以大白猪为研究对象,利用RT-PCR技术克隆SRPK1基因CDS区域,同时利用反向PCR(inverse PCR,I-PCR)技术得到猪部分SRPK1的5′UTR序列;利用生物信息学方法分析SRPK1基因核苷酸序列及编码蛋白序列的结构特点;利用荧光定量PCR(real-time PCR)检测SRPK1在1和30日龄大白猪及杜洛克的心脏、肌肉、脾脏、肝脏、肾脏、肺脏、胃、小肠、大肠、脑的表达情况;构建猪骨骼肌损伤模型,研究在骨骼肌发育过程中SRPK1基因的表达特性。结果,将克隆片段拼接得到长2499bp的大白猪SRPK1基因序列,包含了1968bp完整CDS区域,分析表明其编码含656个氨基酸的蛋白质;包括2个P_Kc结构域,猪SRPK1蛋白序列与人和黑猩猩的相似性较高。通过生物信息学方法预测所得5′UTR含有多个转录结合位点:HSF1、HSF2、Ik-1、IK-2、SRY、SP1MyoD、USF、E47、p300、CP2、RREB-1、E2F和AP-1。利用荧光定量PCR研究发现,表达结果显示猪该基因表达具有组织和种间特异性,主要都是在胃、大肠和小肠中表达。SR-PK1基因在整个损伤修复过程中的表达反复出现升高和降低。5′UTR处有多个和肌肉生长相关蛋白的转录结合位点,主要在消化系统组织表达,在骨骼肌修复中表达上调,推测其可能与骨骼肌细胞发育或其他肌肉生长相关因子的表达有关。 展开更多
关键词 精氨酸/丝氨酸蛋白激酶1 实时荧光定量pcr 反向pcr 骨骼肌损伤模型 生物信息学分析
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Clone and Bioinformatics Analysis of Pig SRPK1 Gene 5' UTR Region
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作者 E Guangxin LIU Di +2 位作者 YANG Xiuqin ZHANG Dongjie YU Cui 《Journal of Northeast Agricultural University(English Edition)》 CAS 2011年第3期22-27,共6页
Part 5' UTR region of pig SRPK1 gene was cloned by inverse PCR (I-PCR), then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool (TFSEARCH) and 32 binding sites of tr... Part 5' UTR region of pig SRPK1 gene was cloned by inverse PCR (I-PCR), then a 425 bp gene sequence was acquired. A promoter region in -1--309 bp was predicted by online tool (TFSEARCH) and 32 binding sites of transcription with scores higher than 85 were getten, of which scores of Spl, MyoD, and HSF2 were over 90. Some of these binding sites of transcription factors were connected with promoters, but TATA-box, which was important to gene expression, hadn't been found in this region. By using PCR-SSCP method to search SNPs this part (5' UTR of SRPK1 in pig), total of 40 Large White pigs were obtained as the research objects, but no polymorphism were found. Thus, 5' UTR of SRPK1 was speculated with a characteristic of high conservation, while it might have been directly or indirectly selected in commercial breeding. The paper provided a further feature of SRPK1 gene in molecular genetics. 展开更多
关键词 PiG SRPK1 BiOiNFORMATiCS transcription factor SSCP inverse pcr i-pcr
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