猪MITF-M的转录调控分析

doi:10.11843/j.issn.0366-6964.2018.11.006

Abstract

Abstract:

This experiment was conducted to study the transcription regulation region activity of pig MITF-M (microphthalmia associated transcription factor M) gene and verify the important transcription factors. PCR amplification with pig DNA as a template was conducted to get the transcription regulation region of pig MITF-M, and 5'-end deletion recombinant vector were obtained by gene cloning. The transcriptional activity of MITF-M was tested via dual luciferase reporter gene experiment, and RNA interference were carried out to verify important transcription factors. The results showed that a 6 414 bp transcription regulation region was cloned; Nine 5'-end deletion recombinant vectors were constructed; Three important regulatory regions of MITF-M were identified (P < 0.001),locating in —1 140-—1 397,—5 429-—6 036 bp positive regulation regions and —2 470-—3 753 negative regulation region; The transcription factor KLF4 significantly negatively regulated the transcription activity of MITF-M(P<0.01). These results indicated that the KLF4 inhibited the transcriptional activity of MITF-M and regulated melanin synthesis and deposition in the pigment cells and affected the pig coat color. These data laid the foundation for further studying the expression mechanism of pig MITF-M, and was of great significance for studying the regulation mechanism of pigs coat color.

Key words: pig, MITF-M, transcriptional activity, KLF4

CLC Number:

S828.2

ZHANG Ting-huan, CHAI Jie, CHEN Lei, LONG Xi. Transcriptional Regulation Analysis of Pig MITF-M Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(11): 2349-2358.

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Transcriptional Regulation Analysis of Pig MITF-M Gene

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