Sequence Characteristics,Tissue Expression and Effect on Milk Fat Synthesis of SIRT7 Gene in Dairy Goats

doi:10.7668/hbnxb.20195461

Abstract

Abstract:

The aim of this study was to clone the silent mating type information regulator 2-related enzyme 7(SIRT7)gene,detect its expression pattern in different tissues and investigate its effect on milk fat synthesis in mammary epithelial cells of dairy goats.The results showed that by designing the primers of SIRT7 gene,the sequence of SIRT7 gene in dairy goat was successfully cloned,with 1 376 bp and a coding region of 1 203 bp,encoding 400 amino acids and showing the highest homology with sheep.SIRT7 was a hydrophilic protein,free of signal peptides and transmembrane structures,with secondary structures such as irregular coils,α-helices,extended chains and β-turns.The relative expression of SIRT7 gene was the highest in the liver tissues of dairy goats,and it also had certain expression levels in other tissues by qRT-PCR.The pcDNA3.1-SIRT7 overexpression vector was successfully constructed and transfected into mammary epithelial cells of dairy goats.Compared with the control group,the mRNA level of SIRT7 gene in the overexpression group was up-regulated about 33 times.After SIRT7 gene overexpression,the expression of stearoyl-CoA desaturase-1(SCD1)gene was significantly decreased,and the expression levels of cluster of differentiation 36(CD36),diacylglycerol acyltransferase 1(DGAT1),sterol regulatory element binding protein 1(SREBP1)genes extremely significantly reduced.The content of triglyceride in cells was significantly decreased.The results of oil red O staining showed that the accumulation of lipid droplets around the nucleus was significantly reduced.Thus, SIRT7 gene inhibited lipid metabolism in mammary epithelial cells of dairy goats,which laid a foundation for further exploration of the regulatory mechanism of SIRT7 in mammary epithelial cells of dairy goats.

Key words: Dairy goat, SIRT7, Gene cloning, Tissue expression, Milk fat synthesis

CLC Number:

S827

LI Zhongying, GUO Zhennan, ZHU Honglin, LI Jialin, SHI Huibin, HAN Haoyuan, LI Jun, QUAN Kai. Sequence Characteristics,Tissue Expression and Effect on Milk Fat Synthesis of SIRT7 Gene in Dairy Goats[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(3): 199-206. doi: 10.7668/hbnxb.20195461.

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Figures/Tables 15

References 26

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基因 Gene	引物序列(5'—3') Primer sequence (5'—3')	退火温度/℃ Annealing temperature	用途 Application
SIRT7	F:GTCTCCTCAGGAGCGATG	53.5	PCR
	R:CACGAGGACAAATGGCTA
SIRT7	F:GATTACCGGGGCCCTAATGG	60.0	qRT-PCR
	R:TCGCAGTTCTGAGACACCAC
SREBP1	F:ACGCCATCGAGAAACGCTAC	60.0	qRT-PCR
	R:GTGCGCAGACTCAGGTTCTC
SCD1	F:CCATCGCCTGTGGAGTCAC	60.0	qRT-PCR
	R:GTCGGATAAATCTAGCGTAGCA
FADS2	F:GGGCTCCACCACCGTGTTCCA	60.0	qRT-PCR
	R:GCTCTGCTGGGCCTGCAGCTG
CD36	F:GTACAGATGCAGCCTCATTTCC	60.0	qRT-PCR
	R:TGGACCTGCAAATATCAGAGGA
DGAT1	F:AAGAGGCTCACCCTGTCGCA	60.0	qRT-PCR
	R:ACTGTTGTTGTCCATGGACG
ATGL	F:GGAGCTTATCCAGGCCAATG	60.0	qRT-PCR
	R:TGCGGGCAGATGTCACTCT
UXT	F:TGTGGCCCTTGGATATGGTT	60.0	qRT-PCR
	R:GGTTGTCGCTGAGCTCTGTG

基因 Gene	引物序列(5'—3') Primer sequence (5'—3')	退火温度/℃ Annealing temperature	用途 Application
SIRT7	F:GTCTCCTCAGGAGCGATG	53.5	PCR
	R:CACGAGGACAAATGGCTA
SIRT7	F:GATTACCGGGGCCCTAATGG	60.0	qRT-PCR
	R:TCGCAGTTCTGAGACACCAC
SREBP1	F:ACGCCATCGAGAAACGCTAC	60.0	qRT-PCR
	R:GTGCGCAGACTCAGGTTCTC
SCD1	F:CCATCGCCTGTGGAGTCAC	60.0	qRT-PCR
	R:GTCGGATAAATCTAGCGTAGCA
FADS2	F:GGGCTCCACCACCGTGTTCCA	60.0	qRT-PCR
	R:GCTCTGCTGGGCCTGCAGCTG
CD36	F:GTACAGATGCAGCCTCATTTCC	60.0	qRT-PCR
	R:TGGACCTGCAAATATCAGAGGA
DGAT1	F:AAGAGGCTCACCCTGTCGCA	60.0	qRT-PCR
	R:ACTGTTGTTGTCCATGGACG
ATGL	F:GGAGCTTATCCAGGCCAATG	60.0	qRT-PCR
	R:TGCGGGCAGATGTCACTCT
UXT	F:TGTGGCCCTTGGATATGGTT	60.0	qRT-PCR
	R:GGTTGTCGCTGAGCTCTGTG

分析工具 Analysis tool	网址 Website	功能 Function
Blast	https: //blast.ncbi.nlm.nih.gov/Blast.cgi	同源性比对分析
ORF Finder	https: //www.ncbi.nlm.nih.gov/orffinder/	开放阅读框分析
ExPASy	https://www.expasy.org	蛋白质理化性质分析
ProScale	https: //web.expasy.org/protscale	蛋白质亲疏水性分析
SignalP	http: //www.cbs.dtu.dk/services/SignalP/	蛋白质信号肽分析
TMHMM 2.0	https://services.healthtech.dtu.dk/service.php?TMHMM-2.0	蛋白质跨膜结构预测
SOPMA	https://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html	蛋白质二级结构预测
SWISS-MODEL	http: //swissmodel.expasy.org/	三级结构预测

分析工具 Analysis tool	网址 Website	功能 Function
Blast	https: //blast.ncbi.nlm.nih.gov/Blast.cgi	同源性比对分析
ORF Finder	https: //www.ncbi.nlm.nih.gov/orffinder/	开放阅读框分析
ExPASy	https://www.expasy.org	蛋白质理化性质分析
ProScale	https: //web.expasy.org/protscale	蛋白质亲疏水性分析
SignalP	http: //www.cbs.dtu.dk/services/SignalP/	蛋白质信号肽分析
TMHMM 2.0	https://services.healthtech.dtu.dk/service.php?TMHMM-2.0	蛋白质跨膜结构预测
SOPMA	https://npsa-pbil.ibcp.fr/cgi-bin/npsa_automat.pl?page=npsa_sopma.html	蛋白质二级结构预测
SWISS-MODEL	http: //swissmodel.expasy.org/	三级结构预测

氨基酸 Amino acid	数量/个 Amount	含量/% Content	氨基酸 Amino acid	数量/个 Amount	含量/% Content
Arg	46	11.5	Gln	17	4.2
Leu	44	11.0	Asp	16	4.0
Ala	32	8.0	His	12	3.0
Glu	32	8.0	Cys	11	2.8
Gly	31	7.8	Ile	11	2.8
Ser	29	7.2	Tyr	8	2.0
Thr	23	5.8	Asn	7	1.8
Lys	22	5.5	Met	7	1.8
Val	22	5.5	Trp	7	1.8
Pro	19	4.8	Phe	4	1.0

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