摘要
本实验以扬州鹅为素材,对鹅胰岛素样生长因子-Ⅰ(IGF-Ⅰ)基因组部分序列进行了克隆、测序,并用PCR-SSCP的方法对其多态性进行了检测。主要结果如下:
1.得到了鹅IGF-Ⅰ基因的5’调控区、外显子1、4及内含子1、3的全部或部分序列,所测序得到的内含子1、内含子3和外显子4序列已上传至Genbank,序列号分别为EU072031、EU111743。本研究首次得到了鹅IGF-Ⅰ基因的内含子1和3序列。序列比对发现,鹅IGF-Ⅰ基因与鸡、鸭同源性较高,无论是外显子还是内含子都大于80%(除内含子3),外显子1与鸡、鹌鹑、火鸡、鸭、鸵鸟、鹅等禽类同源性在98%以上,与梅花鹿、山羊、猪、牛、北极狐、绵羊、小鼠和人等哺乳动物同源性在89%~94%之间,与乌颊鱼、罗非鱼的同源性在60%~65%之间。对鹅5’调控区和外显子1序列进行转录因子结合位点和启动子区域预测,发现有多个转录因子结合位点和一个启动子区域。进化分析表明IGF-Ⅰ基因的进化与物种进化相一致,反映了IGF-Ⅰ基因在进化中的保守性及生理功能的重要性。
2.根据鹅IGF-Ⅰ基因的5’调控区、外显子1和内含子1部分序列设计了7对引物,用PCR-SSCP的方法对其进行单核苷酸多态性(SNP)检测。结果未发现多态性。
In this study, the partial genomic sequences of Insulin-like Growth Factor-ⅠGene (IGF-Ⅰ) of yangzhou goose were cloned and sequenced, and their DNA polymorphism was detected by PCR-SSCP. Main results are as follows:
1. Complete sequences of exon 1, intron 1 and partial sequences of 5’-UTR, intron 3 and exon 4 were obtained in the study. The sequences of intron 1, intron 3 and exon 4 were submitted to Genbank, with accession NO. EU111743 and EU072031. The sequences of intron 1 and 3 of goose IGF-Ⅰgene were reported for the first time. By analysis of homology, the homology of chicken, duck and goose is more than 80% (except intron 3). And the homology of chicken, quail, turkey, duck, ostrich and goose IGF-Ⅰexon 1 is above 98%. The homology between exon 1 and sika deer, goat, pig, cattle, arctic fox, sheep, mouse and human is between 89% and 94%. And the homology between exon 1 and gilthead, tilapia is between 60% and 65%. Several transcription factor binding sites and one promoter was found in 5’–UTR and exon 1 after prediction. The evolution analysis indicated the high identities both in species and IGF-Ⅰgene. The gene was conservation in the process of evolution and had important physiological function.
2. Single nucleotide polymorphism (SNP) of IGF-Ⅰgene were detected with seven pairs of primers by PCR-SSCP in 5’-UTR, exon 1 and intron 1 of IGF-Ⅰgene in yangzhou goose. No single nucleotide polymorphism was found.
引文
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