Goat MZB1 Promotes Intramuscular Adipocyte Differentiation

doi:10.7668/hbnxb.20195540

Abstract

Abstract:

In order to clarify the regulatory role of marginal zone B and B1 cell specific protein(MZB1)in intramuscular adipose differentiation of goats,the study first cloned the full-lenth sequence of goat MZB1 gene and analyzed the sequence to obtain the relevant biological characteristics;then used Real-time Quantitative PCR(qPCR)to construct the expression profile of goat MZB1 in tissues and cells;finally,the goat MZB1 overexpression plasmid was transfected into the target cells,followed by differentiation induction;the effects of MZB1 overexpression on the differentiation process were evaluated through morphological analysis and qPCR.The results showed that the goat MZB1 gene sequence was 860 bp in length,including 570 bp CDS region,encoding 189 amino acids;goat MZB1 protein was not only acidic but also negatively charged,which could be stably existed in the cells.Goat MZB1 expression was detected in eight tissues,including the heart and the liver,and the highest expression was found in the spleen.The expression of MZB1 in goat intramuscular adipocytes after 72 h of oleic acid-induced differentiation was significantly higher than that before oleic acid-induced differentiation.Goat MZB1 overexpression increased the number and size of lipid droplets in goat intramuscular adipocytes;the expression levels of FASN,PPARγ,C/EBPα,SREBP1,and C/EBPβ were highly significantly upregulated.The above results suggest that MZB1 plays a positive regulatory role in goat intramuscular adipocyte differentiation,and this positive regulation may be achieved by increasing the expression of the adipose differentiation marker genes PPARγ,C/EBPα,C/EBPβ,and the fatty acid synthesis marker genes SREBP1 and FASN.

Key words: Goat, MZB1, Intramuscular fat cells, Overexpression, Cell differentiation

CLC Number:

S827

DONG Yaohui, WANG Yong, LI Yanyan, WANG Jinling, WANG Youli, LIU Wei, XING Jiani, LIN Yaqiu. Goat MZB1 Promotes Intramuscular Adipocyte Differentiation[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(5): 216-223. doi: 10.7668/hbnxb.20195540.

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Figures/Tables 9

References 28

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基因 Gene	引物序列(5'—3') Primer sequence(5'—3')	Tm/℃	产物长度/bp Product length	用途 Application
MZB1	S:CAGAGAGGAACCCACACACC	60	707	PCR
	A:CAGAAAGGAGGGAGGCAGAT
MZB1	S:CTATGGAGTTGGAGAAGTGGAC	58	261	qPCR
	A:TCTGTCCTTGTGTCTGGAACCT
UXT	S: ATGGAGTCCTTGGTGAGGTTGT	60	182	内参
	A:GCAAGTGGATTTGGGCTGTAAC
C/EBPα	S: AGGCGGTCATTGTCACTGGT	58	142	qPCR
	A: CCGTGGACAAGAACAGCAAC
PPARγ	S:AAGCGTCAGGGTTCCACTATG	60	197	qPCR
	A:GAACCTGATGGCGTTATGAGAC
C/EBPβ	S:CAAGAAGACGGTGGACAAGC	66	204	qPCR
	A:AACAAGTTCCGCAGGGTG
LPL	S:TCCTGGAGTGACGGAATCTGT	60	174	qPCR
	A:GACAGCCAGTCCACCACGAT
SREBP1	S:AGTGGTGGGCCTCTCTGA	58	127	qPCR
	A:GCAGGGGTTTCTCGGACT

基因 Gene	引物序列(5'—3') Primer sequence(5'—3')	Tm/℃	产物长度/bp Product length	用途 Application
MZB1	S:CAGAGAGGAACCCACACACC	60	707	PCR
	A:CAGAAAGGAGGGAGGCAGAT
MZB1	S:CTATGGAGTTGGAGAAGTGGAC	58	261	qPCR
	A:TCTGTCCTTGTGTCTGGAACCT
UXT	S: ATGGAGTCCTTGGTGAGGTTGT	60	182	内参
	A:GCAAGTGGATTTGGGCTGTAAC
C/EBPα	S: AGGCGGTCATTGTCACTGGT	58	142	qPCR
	A: CCGTGGACAAGAACAGCAAC
PPARγ	S:AAGCGTCAGGGTTCCACTATG	60	197	qPCR
	A:GAACCTGATGGCGTTATGAGAC
C/EBPβ	S:CAAGAAGACGGTGGACAAGC	66	204	qPCR
	A:AACAAGTTCCGCAGGGTG
LPL	S:TCCTGGAGTGACGGAATCTGT	60	174	qPCR
	A:GACAGCCAGTCCACCACGAT
SREBP1	S:AGTGGTGGGCCTCTCTGA	58	127	qPCR
	A:GCAGGGGTTTCTCGGACT

分析内容 Analysis content	工具(网址) Tool(Website)
寻找开放阅读框(ORF)Finding open reading frame (ORF)	ORF finder(https://www.bioinformatics.org/sms2/orf_find.html)
获取氨基酸序列Amino acid sequence acquisition	(https://www.ncbi.nlm.nih.gov/orffinder/)
信号肽位点预测Signal peptide site prediction	SignalP-5.0 Server(https://services.healthtech.dtu.dk/services/SignalP-5.0/)
蛋白质理化性质分析	Protparam(https://web.expasy.org/protparam/)
Analysis of physical and chemical properties of proteins
跨膜结构域预测Transmembrane domain prediction	TMHMM 2.0(https://services.healthtech.dtu.dk/services/TMHMM-2.0/)
功能结构域预测Functional domain prediction	Conserved Domain(https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi)
磷酸化、糖基化位点预测	Netphos、NetOGlyc、NetNGlyc(https://services.healthtech.dtu.dk/services/NetPhos-3.1/、
Prediction of phosphorylation and glycosylation sites	https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/、 https://services.
	healthtech.dtu.dk/services/NetPhos-3.1/)
蛋白质二级结构预测Protein secondary structure prediction	SOPMA(https://npsa.lyon.inserm.fr/cgibin/npsa_automat.pl?page=/NPSA/npsa_sopma.html)
相互作用蛋白质预测Interaction protein prediction	STRING数据库(https://cn.string-db.org/)
蛋白质三级结构预测Tertiary structure prediction of proteins	SWISS-MODEL(https://swissmodel.expasy.org/)

分析内容 Analysis content	工具(网址) Tool(Website)
寻找开放阅读框(ORF)Finding open reading frame (ORF)	ORF finder(https://www.bioinformatics.org/sms2/orf_find.html)
获取氨基酸序列Amino acid sequence acquisition	(https://www.ncbi.nlm.nih.gov/orffinder/)
信号肽位点预测Signal peptide site prediction	SignalP-5.0 Server(https://services.healthtech.dtu.dk/services/SignalP-5.0/)
蛋白质理化性质分析	Protparam(https://web.expasy.org/protparam/)
Analysis of physical and chemical properties of proteins
跨膜结构域预测Transmembrane domain prediction	TMHMM 2.0(https://services.healthtech.dtu.dk/services/TMHMM-2.0/)
功能结构域预测Functional domain prediction	Conserved Domain(https://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi)
磷酸化、糖基化位点预测	Netphos、NetOGlyc、NetNGlyc(https://services.healthtech.dtu.dk/services/NetPhos-3.1/、
Prediction of phosphorylation and glycosylation sites	https://services.healthtech.dtu.dk/services/NetNGlyc-1.0/、 https://services.
	healthtech.dtu.dk/services/NetPhos-3.1/)
蛋白质二级结构预测Protein secondary structure prediction	SOPMA(https://npsa.lyon.inserm.fr/cgibin/npsa_automat.pl?page=/NPSA/npsa_sopma.html)
相互作用蛋白质预测Interaction protein prediction	STRING数据库(https://cn.string-db.org/)
蛋白质三级结构预测Tertiary structure prediction of proteins	SWISS-MODEL(https://swissmodel.expasy.org/)

氨基酸 Amino acid	数目/个 Amount	百分比/% Percentage	氨基酸 Amino acid	数目/个 Amount	百分比/% Percentage
Ala	14	7.4	Leu	27	14.3
Arg	10	5.3	Lys	6	3.2
Asn	0	0.0	Met	7	3.7
Asp	10	5.3	Phe	3	1.6
Cys	6	3.2	Pro	11	5.8
Gln	10	1.7	Ser	12	6.3
Glu	15	7.9	Thr	12	6.3
Gly	20	10.6	Trp	4	2.1
His	5	2.6	Tyr	5	2.6
Ile	4	2.1	Val	8	4.2
Pyl	0	0.0	Sec	0	0.0

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