Characteristics and Expression Profile of KRT71 Screened by Suppression Subtractive Hybridization cDNA Library in Curly Fleece Chinese Tan Sheep

DNA and Cell Biology
Xiaolong KangMeiying Fang

Abstract

As an important commercial trait for sheep, curly fleece has a great economic impact on production costs and efficiency in sheep industry. To identify genes that are important for curly fleece formation in mammals, a suppression subtractive hybridization analysis was performed on the shoulder skin tissues exposed to two different growth stages of Chinese Tan sheep with different phenotypes (curly fleece and noncurling fleece). BLAST analysis identified 67 differentially expressed genes, of which 31 were expressed lower and 36 were expressed higher in lambs than in adult sheep. Differential expressions of seven randomly selected genes were verified using quantitative real-time polymerase chain reaction (qRT-PCR). KRT71 gene was selected for further study due to its high correlation with the curly hair phenotype in various mammal species. Semi-qPCR showed distinctively high expression of KRT71 in skin tissues. Moreover, qPCR result showed a significantly higher expression of KRT71 in curly fleece than noncurling Tan sheep. The luciferase assay and electrophoresis mobility shift assay showed that there were transcription factor binding sites in the promoter region of KRT71 related to the differential expression of KRT71 at the two...Continue Reading

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Datasets Mentioned

BETA
BC103917.1
BC125348.1
JX975451

Methods Mentioned

BETA
electrophoresis
PCR
transfection
Assay
electrophoretic mobility shift assay

Software Mentioned

Secondary Structure Prediction Method ( SOPMA
Ensembl
_ Check
Primer5
Primer Premier
PolyA
KOBAS
BLASTN
CAP3
TFSEARCH

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