Cloning and Polymorphism Analysis of VIPR-1 Gene in Black Muscovy Duck
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摘要: VIPR-1基因是研究家禽就巢性状的重要候选基因,VIPR-1基因的克隆和测序能为进一步研究黑番鸭就巢性状的单核苷酸多态性奠定理论基础,为鸭分子遗传育种提供基因素材。本研究以黑番鸭基因组为模板进行PCR扩增,获取黑番鸭VIPR-1基因序列[1 864bp(序列1)和1 673bp(序列2)的基因片段],并进行目的基因片段克隆和测序。结果分析表明,序列1包含第5外显子(101bp)、第5内含子(1 630bp)及第6外显子(133bp)的完整序列;序列2包含第12外显子(42bp)、第12内含子(1 455bp)的完整序列和第13外显子(176bp)的部分序列。根据基因序列特征,对获取的2段黑番鸭VIPR-1基因序列分别设计4对引物,进行扩增序列测序比对。结果发现:在序列1第318处、454处存在C/T突变,第547处存在A/G突变,第923处存在A/T突变;序列2第89处、944处存在C/T突变,第662处存在G/A突变,第1 031处存在A/G突变,第1 334处存在A/C突变。黑番鸭VIPR-1基因序列的克隆与单核苷酸多态性SNP突变位点的发现为进一步开展VIPR-1基因的多态性与黑番鸭就巢性状相关研究奠定了基础。Abstract: As a critical candidate gene of poultry broodiness,the length of 1 864 bp(target 1) and 1 673 bp(target 2) gene fragments of the vasoactive intestinal peptide-1 gene were amplified with polymerase chain reaction method from the genomic DNAs Abstracted from the blood of black Muscovy duck.The results of sequence analysis demonstrated that the target fragment 1 included the complete sequence of the 5th exon(101 bp),the 5th intron(1 630 bp) and the 6th exon(133 bp).The target fragment 2 contained the complete sequence of the 12th exons(42 bp),the 12th introns(1 455 bp) and partial sequence of the 13th exons(176 bp).Another four pairs of specific primers(P1-P8) were designed according to the two sequences fragments,respectively.The results showed that the target fragment 1 had C/T mutation at position 318 bp and 454 bp,A/G mutation at 547 bp,A/T mutation at 923 bp;the target fragment 2 had C/T mutation at 89 bp and 944 bp,A/G mutation at 662 bp and 1 031 bp,A/C mutation at 1 334 bp.The characterization and mutations discovery with the VIPR-1 gene provided the basic evidences for the further study on the association of its polymorphisms with broodiness trait with the black Muscovy ducks.
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Key words:
- black muscovy duck /
- VIPR-1 gene /
- clone /
- polymorphism analysis
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