畜牧兽医学报
畜牧獸醫學報
축목수의학보
2009年
11期
1600-1608
,共9页
陈书明%晋大鹏%王立贤%张龙超%颜华%程笃学
陳書明%晉大鵬%王立賢%張龍超%顏華%程篤學
진서명%진대붕%왕립현%장룡초%안화%정독학
猪SP-A基因%启动子区%克隆%染色体步移%序列分析
豬SP-A基因%啟動子區%剋隆%染色體步移%序列分析
저SP-A기인%계동자구%극륭%염색체보이%서렬분석
porcine surfactant protein A gene%promoter region%clone%Genome Walking%sequence analysis
为了探明猪SP-A基因的启动子及其转录调控机制,设计6条特异性引物,采用染色体步移、克隆测序以及序列比对分析,从大白猪基因组中扩增出一段长度为1 033 bp的猪SP-A基因的上游序列(DQ985806),其GC含量约为55%.经过生物信息学分析,确定了其转录起始位点及活性区域;发现在转录起始位点上游-33 bp处存在1个TATA-box,另外还发现了GC-box、CAAT-box、GT-box以及转录起始子;该序列还包含Spl、TF和NF-κB等转录因子结合位点以及1个特殊重复序列5'-CATCACTGT-3'.上述试验结果表明,所克隆的1 033 bp的DNA序列是大白猪SP-A基因的启动子区序列.
為瞭探明豬SP-A基因的啟動子及其轉錄調控機製,設計6條特異性引物,採用染色體步移、剋隆測序以及序列比對分析,從大白豬基因組中擴增齣一段長度為1 033 bp的豬SP-A基因的上遊序列(DQ985806),其GC含量約為55%.經過生物信息學分析,確定瞭其轉錄起始位點及活性區域;髮現在轉錄起始位點上遊-33 bp處存在1箇TATA-box,另外還髮現瞭GC-box、CAAT-box、GT-box以及轉錄起始子;該序列還包含Spl、TF和NF-κB等轉錄因子結閤位點以及1箇特殊重複序列5'-CATCACTGT-3'.上述試驗結果錶明,所剋隆的1 033 bp的DNA序列是大白豬SP-A基因的啟動子區序列.
위료탐명저SP-A기인적계동자급기전록조공궤제,설계6조특이성인물,채용염색체보이、극륭측서이급서렬비대분석,종대백저기인조중확증출일단장도위1 033 bp적저SP-A기인적상유서렬(DQ985806),기GC함량약위55%.경과생물신식학분석,학정료기전록기시위점급활성구역;발현재전록기시위점상유-33 bp처존재1개TATA-box,령외환발현료GC-box、CAAT-box、GT-box이급전록기시자;해서렬환포함Spl、TF화NF-κB등전록인자결합위점이급1개특수중복서렬5'-CATCACTGT-3'.상술시험결과표명,소극륭적1 033 bp적DNA서렬시대백저SP-A기인적계동자구서렬.
In order to explore the mechanism of porcine SP-A gene transcription regulation by studying the promoter region of porcine SP-A gene, the upstream sequence of Large White por-cine SP-A gene was attained by Genome Walking PCR,sequencing and sequences alignment anal-ysis. The DNA sequence of 1 033 bp (DQ985806) was amplified. The ratio of G and C in the se-quence was about 55%. The sequence analysis showed that there were a TATA-box at-33 bp and GC-box, CAAT-box, GT-box and transcriptional initiator. Meanwhile, some transcriptional factor binding sites including Spl,TF,NF-κB,and so on were detected. In addition,a special re-petitive sequence of 5'-CATCACTGT-3' was also detected. The results showed that the cloned sequence (1 033 bp) may be considered as the promoter region of Large White porcine SP-A gene.