| Fineness of cashmere is the key to evaluate the quality and price of cashmere,and the nature of its phenotypic characteristics is determined by gene expression.In recent years,more and more studies have been conducted on cashmere related non-coding RNA(nc RNA).However,studies on CircRNA and cashmere fineness regulation are limited.CircRNA is an endogenous non-coding RNA(nc RNA)with covalently closed ring structure.CircRNA has a unique circular structure,It is more stable than m RNA,and has a longer half-life.Because CircRNA is a closed ring structure it is very stable in most eukaryotes.We collected skin samples of Liaoning cashmere goat(LCG)and Inner Mongolia cashmere goat(MCG).Different expression of circrna was sequenced.There were 17 up-regulated and 15 downregulated circ RNAs in LCG compared with MCG skin.In order to find the differentially expressed circrna in LCG,We verified 10 circrnas by q PCR,Four circrnas: circna128,circrna6854,circrna4154 and circrna3620 were highly expressed in Liaoning cashmere goats.In addition,the regulatory network of circrna mi RNA is bioinformatic,which may help to understand the molecular mechanism involved in the regulation of cashmere fineness.1.Analysis of cashmere fineness related circ RNA sequencing resultsIn this study,a total of 13320 circrnas were identified from the skin tissue RNA SEQ of Liaoning Cashmere Goat and Inner Mongolia cashmere goat,It includes 610 intronic RNA(ci RNA s).7531 and 8943 circular RNAs were detected in LCG and MCG libraries,respectively.In LCG and MCG,the amount of circ RNA in the range of 200-400 BP was the most.Most of the circular RNAs contain 2-7 exons.We confirmed the differential expression of circrna in coarse and fine LCG skin by q PCR.The results of RNA SEQ showed that the expression level of cirna128 was the highest in the up-regulated circ RNA.,The expression level of circrna6854 was the highest in the down-regulated circ RNA.The results of circ RNA128,circ RNA6854,circ RNA3620 and circ RNA4154 were significantly different in RNA SEQ and q PCR.This suggests that they may play an active role in cashmere goats with different fiber diameters.At the same time,we found a highly expressed host gene TCHH.2.Ce RNA regulatory network of cashmere fineness key gene TCHHWe predicted the regulatory network of TCHH gene by Ce RNA prediction.TCHH gene is the target gene of five mi RNAs.Subsequently,five mi RNAs were detected by fluorescence quantitative PCR.The results showed that mi R-126 was highly expressed in fine type of Liaoning cashmere goat;MSTRG-5429.1 was highly expressed in the coarse type of Liaoning cashmere goat,It may play a positive role in regulating the cashmere quality of Liaoning cashmere goats.3.SNP detection of TCHH gene and its correlation with cashmere finenessIn order to understand the effect of gene TCHH on cashmere yield and fineness.Blood samples of 300 Liaoning cashmere goats were collected and sequenced,Four SNP mutation sites were screened in the exon of gene TCHH,Among them,G112 A,G287T and A555 G mutations produced three genotypes,The A585 T mutation produced two genotypes.The results showed that G112 A,G287T and A585 T genotypes had negative effects on cashmere yield,The yield of cashmere decreased to some extent,The GG genotype mutation of A555 G locus can significantly improve cashmere yield of cashmere goats(4.5%)。... |
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