HGTP基因与羊毛细度的相关性分析及其启动子活性研究

doi:10.16431/j.cnki.1671-7236.2019.10.020

Abstract

Abstract: This study was aimed to construct a high glycine-tyrosine protein (HGTP) promoter luciferase reporter vector,verify its activity,and elucidate the relationship between the expression difference of HGTP gene in the skin and hair follicle tissues of Suffolk and Chinese Merino sheep and the economic traits of wool.The HGTP gene promoter fragment was amplified by PCR and cloned into pGL3-Basic vector to construct a recombinant plasmid,which was verified by double enzyme digestion and nucleic acid sequence identification.Skin fibroblasts were transfected and the activity of the reporter gene was detected.The fiber diameter and length of the wool were measured,and the expression of HGTP gene in the hair follicle tissues of different breeds of sheep were analyzed by Real-time PCR.The results showed that the differences between wool fiber diameter and natural length of wool in Suffolk and Chinese Merino sheep were extremely significant (P<0.01).There was a highly significant positive correlation between the expression of HGTP gene and wool fiber diameter (P<0.01),a significant medium negative with the natural length of wool (P<0.05),and there was a very significant high negative correlation between wool diameter and natural length (P<0.01).The expression of HGTP gene in skin tissues of Suffolk and Chinese Merino sheep were extremely significant (P<0.01).The promoter activity luciferase assay showed that the KAP6.1,KAP7 and KAP8.1 promoter elements were expressed in both sheep skin fibroblasts and 3T3 cells,and the activity of the three promoter elements in sheep fibroblasts were extremely significantly higher than that in 3T3 cells (P<0.01).HGTP gene could be used as a candidate gene for studying the diameter and natural length of wool fibers.Expression vectors of different lengths were constructed and transfected into skin fibroblasts,both promoter elements were obtained to drive the expression of foreign genes at the cellular level in vitro.The functional study of HGTP gene promoter in sheep provided a theoretical basis for regulating wool development.

Key words: sheep; high glycine-tyrosine protein (HGTP); promoter; dual luciferase reporter gene

CLC Number:

S813.3

WANG Jingjing, YANG Hanyulu, DAI Rong, YANG Hua, SHI Guoqing, YANG Yonglin, WAN Pengcheng. Study on the Correlation Between HGTP Gene and Wool Fineness and the Activity of HGTP Gene Promoter[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(10): 2988-2997.

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Study on the Correlation Between HGTP Gene and Wool Fineness and the Activity of HGTP Gene Promoter

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