Effect of Nucleotide Sequence Variation of STAT3 on Milk Performance Traits of Sheep

doi:10.7668/hbnxb.20195805

Abstract

Abstract:

To explore the association between STAT3 single nucleotide polymorphism and lactation traits of sheep,414 lactating sheep were selected as test subjects,the expression of STAT3 in seven tissues including mammary gland was detected by RT-qPCR,the single nucleotide polymorphism of STAT3 was detected by sanger sequencing,STAT3 was typed by Penta-primer amplification refractory mutation system PARMS,and the effect of the nucleotide sequence variation of the gene on milk performance traits was analysed.RT-qPCR results revealed that STAT3 expression was highest in lungs,followed by liver and mammary tissues,and lowest in spleen.A total of five novel SNPs,c.22+537 C/T,c.22+658 C/T,c.318+126 C/G,c.417+360 G/A and c.417+395 T/C,were identified in the sheep STAT3,and three genotypes were detected at each SNP locus.Association analysis revealed that ewes with the genotype CC at the c.318+126 C/G locus had a higher milk fat rate than those with GG,and CC-type at the c.22+537 C/T locus,CC-type at the c.22+658 C/T locus,GG-type at the c.417+360 G/A locus,and TT-type at the c.417+395 T/C locus all had higher milk fat percentage and dry matter content.The haplotype combination H1H7 exhibited significant positive effects on milk fat percentage,dry matter content,and ash content.Additionally,the average daily milk yield of individuals carrying the haplotype combination H1H2 was increased by 0.266 kg compared to those carrying H1H7.In conclusion,the five nucleotide variation sites of STAT3 screened in this study may provide a theoretical basis for molecular selection of milk performance traits in milk sheep.

Key words: Milk sheep, STAT3, Nucleotide sequence variation, Milk performance traits, Gene expression

CLC Number:

S826

YANG Pan, ZHEN Huimin, HAO Zhiyun, ZHANG Xiyun, CHE Longjie, LI Mingna, REN Chunyan, WANG Xuanyu, ZHAO Yuan, LIU Yuan, WANG Jiqing, WANG Zike. Effect of Nucleotide Sequence Variation of STAT3 on Milk Performance Traits of Sheep[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(5): 198-207. doi: 10.7668/hbnxb.20195805.

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References 37

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引物 Primer	引物序列(5'—3') Primer sequence(5'—3')	扩增区域 Application range	退火温度/℃ Annealing temperature	目的 Purpose
P1	F: CCAGAGCATTGTGAGTGAGC	第8外显子	60	SNP筛选
	R: TCTTCATGCAGCTCTGACCA
P2	F: CTTCCCCTTCTCCAGCTCAT	第9外显子	60	SNP筛选
	R: CAGCTCCACGATTCTCTCCT
P3	F: CTGCCTATTCATCCCTCCCC	第1内含子	62	SNP筛选
	R: GGTGACACGCCTTCCTTTAG
P4	F: TGACCAACAGTATAGCCGCT	第3内含子	62	SNP筛选
	R: CTACGGTGTCCAAGCCTTTG
P5	F: ACTGGCTCCTCTGTCCATTT	第4内含子	60	SNP筛选
	R: TCCCACGCCTAGAAATCTGT
P6	F: GGAAGTGGGGATGGGAAACA	第11内含子	62	SNP筛选
	R: GGAGGAAAAGACCAACCACG
P7	F: TTTGTAGGTAAGGCTGGGCA	3'非翻译区	60	SNP筛选
	R: CCAAGACCAGGTGCGTAAAG
STAT3	F: CGGCCAGCAAAGAATCACAT	编码序列	58	RT-qPCR
	R: CCACAATTCGGGCAATCTCC
β-actin	F: AGCCTTCCTTCCTGGGCATGGA	编码序列	58	RT-qPCR
	R: GGAGCGAAACTGACACCC

引物 Primer	引物序列(5'—3') Primer sequence(5'—3')	扩增区域 Application range	退火温度/℃ Annealing temperature	目的 Purpose
P1	F: CCAGAGCATTGTGAGTGAGC	第8外显子	60	SNP筛选
	R: TCTTCATGCAGCTCTGACCA
P2	F: CTTCCCCTTCTCCAGCTCAT	第9外显子	60	SNP筛选
	R: CAGCTCCACGATTCTCTCCT
P3	F: CTGCCTATTCATCCCTCCCC	第1内含子	62	SNP筛选
	R: GGTGACACGCCTTCCTTTAG
P4	F: TGACCAACAGTATAGCCGCT	第3内含子	62	SNP筛选
	R: CTACGGTGTCCAAGCCTTTG
P5	F: ACTGGCTCCTCTGTCCATTT	第4内含子	60	SNP筛选
	R: TCCCACGCCTAGAAATCTGT
P6	F: GGAAGTGGGGATGGGAAACA	第11内含子	62	SNP筛选
	R: GGAGGAAAAGACCAACCACG
P7	F: TTTGTAGGTAAGGCTGGGCA	3'非翻译区	60	SNP筛选
	R: CCAAGACCAGGTGCGTAAAG
STAT3	F: CGGCCAGCAAAGAATCACAT	编码序列	58	RT-qPCR
	R: CCACAATTCGGGCAATCTCC
β-actin	F: AGCCTTCCTTCCTGGGCATGGA	编码序列	58	RT-qPCR
	R: GGAGCGAAACTGACACCC

位点 Locus	引物序列(5'—3') Primer sequence(5'—3')	备注 Note
SNP1	CCAGAATATCTTTTGGTTGGAAGC	ASP (C)
	TCCAGAATATCTTTTGGTTGGAAGT	ASP (T)
	CTGAATCAGAGTTGCCAAGTGAAC	LSP
SNP2	AGTAATATTCCATTGTCTGTGTGTACC	ASP (C)
	TGAGTAATATTCCATTGTCTGTGTGTACT	ASP (T)
	GTGCTCAAACTTGGGAACAATTAT	LSP
SNP3	AAGGAGAAGTTACCAGCCCTTTC	ASP (C)
	AGGAGAAGTTACCAGCCCTTTG	ASP (G)
	CACATTATTATTTCAATGAACCAACC	LSP
SNP4	CATGATGTAATTAAAAGAATGCAAGAC	ASP (G)
	CATGATGTAATTAAAAGAATGCAAGAT	ASP (A)
	TCATAACTGTCTCCAGGCTCCTA	LSP
SNP5	TTAGCTTTAAACTGCTTGACTTGGA	ASP (T)
	TAGCTTTAAACTGCTTGACTTGGG	ASP (C)
	TAGCCTTTCATAACTGTCTCCAGG	LSP
通用引物1 Universal primer 1	GAAGGTGACCAAGTTCATGCT	CFP (FAM)
通用引物2 Universal primer 2	GAAGGTCGGAGTCAACGGATT	CFP (HEX)

位点 Locus	引物序列(5'—3') Primer sequence(5'—3')	备注 Note
SNP1	CCAGAATATCTTTTGGTTGGAAGC	ASP (C)
	TCCAGAATATCTTTTGGTTGGAAGT	ASP (T)
	CTGAATCAGAGTTGCCAAGTGAAC	LSP
SNP2	AGTAATATTCCATTGTCTGTGTGTACC	ASP (C)
	TGAGTAATATTCCATTGTCTGTGTGTACT	ASP (T)
	GTGCTCAAACTTGGGAACAATTAT	LSP
SNP3	AAGGAGAAGTTACCAGCCCTTTC	ASP (C)
	AGGAGAAGTTACCAGCCCTTTG	ASP (G)
	CACATTATTATTTCAATGAACCAACC	LSP
SNP4	CATGATGTAATTAAAAGAATGCAAGAC	ASP (G)
	CATGATGTAATTAAAAGAATGCAAGAT	ASP (A)
	TCATAACTGTCTCCAGGCTCCTA	LSP
SNP5	TTAGCTTTAAACTGCTTGACTTGGA	ASP (T)
	TAGCTTTAAACTGCTTGACTTGGG	ASP (C)
	TAGCCTTTCATAACTGTCTCCAGG	LSP
通用引物1 Universal primer 1	GAAGGTGACCAAGTTCATGCT	CFP (FAM)
通用引物2 Universal primer 2	GAAGGTCGGAGTCAACGGATT	CFP (HEX)

位点 Locus	群体 Populations	个体数 Number
SNP1			CC	CT	TT	C	T
	H	35	5.71	62.86	31.43	37.14	62.86	0.040 6	0.466 9	1.876 0	0.357 9
	EF	17	11.77	52.94	35.29	38.24	61.76	0.618 2	0.472 3	1.895 1	0.360 8
	F₁	193	8.29	50.78	40.93	33.68	66.32	0.057 6	0.446 7	1.807 4	0.346 9
	F₂	169	10.65	46.75	42.60	34.02	65.98	0.592 1	0.449 0	1.814 7	0.348 2
	平均值		9.11	53.33	37.56	35.77	64.23
SNP2			CC	CT	TT	C	T
	H	35	5.71	34.29	60.00	22.86	77.14	0.869 5	0.352 7	1.544 8	0.290 5
	EF	17	23.53	47.06	29.41	47.06	52.94	0.818 8	0.498 3	1.993 1	0.374 1
	F₁	193	13.47	51.30	35.23	39.12	60.88	0.285 3	0.476 3	1.909 6	0.362 9
	F₂	169	12.43	51.48	36.09	38.17	61.83	0.238 4	0.472 0	1.893 9	0.360 6
	平均值		13.79	46.03	40.18	36.80	63.20
SNP3			CC	CG	GG	C	G
	H	35	14.29	57.14	28.57	42.86	57.14	0.324 1	0.489 8	1.960 0	0.369 8
	EF	17	5.88	47.06	47.06	29.41	70.59	0.582 5	0.415 2	1.710 1	0.329 0
	F₁	193	19.17	52.33	28.50	45.34	54.66	0.438 1	0.495 7	1.982 8	0.372 9
	F₂	169	11.83	55.03	33.14	39.35	60.65	0.046 8	0.477 3	1.913 2	0.363 4
	平均值		12.79	52.89	34.32	39.24	60.76
SNP4			GG	GA	AA	G	A
	H	35	2.86	42.86	54.28	24.29	75.71	0.327 9	0.367 8	1.581 7	0.300 1
	EF	17	17.65	52.94	29.41	44.12	55.88	0.761 3	0.493 1	1.972 7	0.371 5
	F₁	193	11.92	60.10	27.98	41.97	58.03	0.001 2	0.487 1	1.949 7	0.368 5
	F₂	169	12.43	57.99	29.58	41.42	58.58	0.011 3	0.485 3	1.942 8	0.367 5
	平均值		11.22	53.47	35.31	37.95	62.05
SNP5			TT	TC	CC	T	C
	H	35	5.71	40.00	54.29	25.71	74.29	0.780 9	0.382 0	1.618 2	0.309 1
	EF	17	23.53	47.06	29.41	47.06	52.94	0.818 8	0.498 3	1.993 1	0.374 1
	F₁	193	12.95	58.03	29.02	41.97	58.03	0.007 9	0.487 1	1.949 7	0.368 5
	F₂	169	13.02	57.40	29.58	41.72	58.28	0.019 1	0.486 3	1.946 6	0.368 0
	平均值		13.80	50.62	35.58	39.11	60.89

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