FAT1转基因系祖公牛选育及其扩繁研究

doi:10.13304/j.nykjdb.2021.0498

摘要/Abstract

摘要：

n-3多不饱和脂肪酸（n-3 polyunsaturated fatty acids，n-3 PUFAs）是人体必需脂肪酸，对多种疾病有抑制作用。利用超数排卵与胚胎移植技术（superovulation and embryo transfer，MOET）对FAT1转基因牛进行了扩繁，并对F₁代牛进行了生长指标测定与遗传稳定性分析，在此基础上确定了系祖公牛ZK005；用ZK005的冻精经人工授精生产F₂代，并对其进行生长指标测定、遗传稳定性分析和血液生化指标检测。结果显示，原代牛的MOET扩繁效果良好，获得了8头FAT1基因阳性F₁代牛；F₁代牛生长指标与野生型牛无显著差异；外源基因稳定整合，n-3 PUFAs含量显著提高，n-6多不饱和脂肪酸/n-3多不饱和脂肪酸（n-6 polyunsaturated fatty acids/n-3 polyunsaturated fatty acids，n-6/n-3）比值显著降低；F₁代牛各组织中n-3 PUFAs含量显著提高，其中肌肉中提高最明显。从F₁代牛中选择备用系祖公牛ZK005，其精液冷冻后活力达40%，经人工输精获得F₂代牛12头，F₂代牛生长发育与血液生化指标整体上与野生型牛无显著差异。以上研究表明，利用转基因技术选育FAT1转基因牛，可培育富含n-3 PUFAs的转基因牛新品系（种），提高牛肉、奶中n-3 PUFAs含量，且FAT1基因在F₁和F₂代牛中均能稳定有效表达，对转基因牛的健康无不利影响，可为人类提供更营养的食品。

关键词: n-3多不饱和脂肪酸, 系祖, 遗传稳定性, 血液生化指标

Abstract:

n-3 polyunsaturated fatty acids （n-3 PUFAs） are essential fatty acids for human health and can inhibit many diseases. Using transgenic technology to produce FAT1 transgenic cattle can improve the content of n-3 PUFAs in beef and milk， and provide healthy food for human. In this study， after propagation of FAT1 transgenic cattle through superovulation and embryo transfer （MOET） technique， the growth index and genetic stability of F₁ generation cattle were tested. After that， ZK005 bull was determined as ancestry based on the analysis. The F₂ generation was produced by artificial insemination with frozen semen of ZK005， and its growth indexes， genetic stability and blood biochemical indexes were measured. The results showed that 8 FAT1 gene positive F₁ cattle were obtained. There was no significant difference in growth index between F₁ and wild-type cattle. The content of n-3 PUFAs increased significantly and the ratio of n-6/n-3 PUFAs decreased significantly. The content of n-3 PUFAs in various tissues significantly increased， especially in muscles. Total of 12 F₂ generation cattle， were obtained by artificial insemination. There was no significant difference in growth and development and blood biochemical indexes between F₂ generation cattle and wild-type cattle as a whole， while the above research showed that breeding FAT1 transgenic cattle with transgenic technology could cultivate new transgenic cattle strains （species） rich in n-3 PUFAs， improved the content of n-3 PUFAs in beef and milk， and FAT1 gene could be stably and effectively expressed in F₁ and F₂ generation cattle， which had no adverse impact on the health of transgenic cattle and could provide more nutritious food for human beings.

Key words: n-3 polyunsaturated fatty acids, ancestral bull, genetic stability, blood biochemical indexes

中图分类号:

S823

白春玲, 杨磊, 苏广华, 魏著英, 王学侨, 朱琳, 周新宇, 海超, 吴迪, 刘雪霏, 武云喜, 张立, 李光鹏. FAT1转基因系祖公牛选育及其扩繁研究[J]. 中国农业科技导报, 2022, 24(9): 66-78.

Chunling BAI, Lei YANG, Guanghua SU, Zhuying WEI, Xueqiao WANG, Lin ZHU, Xinyu ZHOU, Chao HAI, Di WU, Xuefei LIU, Yunxi WU, Li ZHANG, Guangpeng LI. Breeding and Propagation of FAT1 Transgenic Ancestral Bull[J]. Journal of Agricultural Science and Technology, 2022, 24(9): 66-78.

图/表 10

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类型	编号 Number	目的基因 Target gene	n-6 PUFAs/%	n-3 PUFAs/%	n-6/n-3 PUFAs	n-6/n-3 PUFAs平均值 n-6/n-3 PUFAs average value
FAT1基因阳性牛 FAT1 gene positive cattle	ZK004	+	2.08	1.86	1.12	3.93 b
	ZK005	+	7.69	3.35	2.29
	FD001	+	18.44	8.50	2.17
	FD002	+	37.15	4.65	7.98
	FD003	+	19.77	2.36	8.38
	FD004	+	14.08	5.31	2.65
	FD005	+	12.11	2.93	4.13
	FD006	+	9.86	3.62	2.72
野生型牛 Wild type cattle	WT-1	NA	1.89	0.12	15.75	16.10 a
	WT-2	NA	4.97	0.47	10.57
	WT-3	NA	11.21	0.51	21.98
FAT1基因阴性牛 FAT1 gene negative cattle	Con 1	-	18.11	1.71	10.59	13.14 a
	Con 2	-	4.90	0.38	12.90
	Con 3	-	21.99	1.38	15.93

类型	编号 Number	目的基因 Target gene	n-6 PUFAs/%	n-3 PUFAs/%	n-6/n-3 PUFAs	n-6/n-3 PUFAs平均值 n-6/n-3 PUFAs average value
FAT1基因阳性牛 FAT1 gene positive cattle	ZK004	+	2.08	1.86	1.12	3.93 b
	ZK005	+	7.69	3.35	2.29
	FD001	+	18.44	8.50	2.17
	FD002	+	37.15	4.65	7.98
	FD003	+	19.77	2.36	8.38
	FD004	+	14.08	5.31	2.65
	FD005	+	12.11	2.93	4.13
	FD006	+	9.86	3.62	2.72
野生型牛 Wild type cattle	WT-1	NA	1.89	0.12	15.75	16.10 a
	WT-2	NA	4.97	0.47	10.57
	WT-3	NA	11.21	0.51	21.98
FAT1基因阴性牛 FAT1 gene negative cattle	Con 1	-	18.11	1.71	10.59	13.14 a
	Con 2	-	4.90	0.38	12.90
	Con 3	-	21.99	1.38	15.93

性别 Sex	月龄/月 Month age/mouth	体质量 Body mass/kg		体高 Body height/cm		十字部宽 Height at hip cross/cm		胸围 Bust/cm
性别 Sex	月龄/月 Month age/mouth	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control
母牛Cow	0	28.80	27.48	79.22	79.00	17.18	17.20	72.78	73.04
	3	125.80	115.40	92.22	91.11	24.82	22.88	110.95	110.44
	6	174.45	179.60	99.82	99.74	28.14	27.56	124.90	123.13
	9	218.60	225.15	106.23	106.56	30.45	31.06	136.50	134.70
	12	256.70	248.70	117.06	117.86	35.52	35.45	151.62	153.02
	15	271.20	275.60	122.52	121.79	36.04	36.22	157.53	155.44
	18	373.90	386.25	124.98	125.74	37.84	37.80	167.14	165.13
公牛Bull	0	30.60	28.70	80.00	79.70	17.50	17.40	73.40	73.20
	3	131.40	120.40	92.60	93.05	24.50	25.20	113.25	113.10
	6	201.35	192.20	102.40	102.00	29.20	28.10	126.30	125.45
	9	249.60	248.05	108.15	107.30	32.05	32.20	138.10	137.50
	12	297.10	292.50	118.20	117.80	36.10	35.85	154.00	154.20
	15	332.00	335.40	124.30	123.95	37.50	36.90	159.85	158.10
	18	443.70	456.35	127.20	126.20	39.60	39.50	168.60	168.25

性别 Sex	月龄/月 Month age/mouth	体质量 Body mass/kg		体高 Body height/cm		十字部宽 Height at hip cross/cm		胸围 Bust/cm
性别 Sex	月龄/月 Month age/mouth	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control
母牛Cow	0	28.80	27.48	79.22	79.00	17.18	17.20	72.78	73.04
	3	125.80	115.40	92.22	91.11	24.82	22.88	110.95	110.44
	6	174.45	179.60	99.82	99.74	28.14	27.56	124.90	123.13
	9	218.60	225.15	106.23	106.56	30.45	31.06	136.50	134.70
	12	256.70	248.70	117.06	117.86	35.52	35.45	151.62	153.02
	15	271.20	275.60	122.52	121.79	36.04	36.22	157.53	155.44
	18	373.90	386.25	124.98	125.74	37.84	37.80	167.14	165.13
公牛Bull	0	30.60	28.70	80.00	79.70	17.50	17.40	73.40	73.20
	3	131.40	120.40	92.60	93.05	24.50	25.20	113.25	113.10
	6	201.35	192.20	102.40	102.00	29.20	28.10	126.30	125.45
	9	249.60	248.05	108.15	107.30	32.05	32.20	138.10	137.50
	12	297.10	292.50	118.20	117.80	36.10	35.85	154.00	154.20
	15	332.00	335.40	124.30	123.95	37.50	36.90	159.85	158.10
	18	443.70	456.35	127.20	126.20	39.60	39.50	168.60	168.25

性别 Sex	月龄/月 Month age/month	体质量 Body mass/kg		体高 Body height/cm		十字部宽 Height at hip cross/cm		胸围 Bust/cm
性别 Sex	月龄/月 Month age/month	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control	转基因组 Transgenic	对照组 Control
母牛 Cow	0	33.10	32.41	73.90	72.47	20.90	21.33	76.21	77.53
	3	92.96	90.40	90.70	89.42	23.89	24.26	108.40	107.70
	6	154.10	152.50	99.90	99.50	28.02	28.33	125.90	124.80
	9	185.90	184.00	106.80	104.50	32.41	32.90	137.30	136.10
	12	231.20	229.30	114.70	113.20	35.66	34.45	150.40	147.20
	15	266.60	260.80	117.70	116.20	36.42	36.82	158.50	153.90
	18	310.90	299.80	120.20	119.20	37.92	37.04	169.20	165.70
公牛Bull	0	32.52	31.88	74.40	74.01	21.03	20.44	74.67	72.23
	3	99.70	100.90	93.18	92.69	26.78	26.34	118.30	116.50
	6	170.30	169.40	100.07	101.77	29.01	28.46	129.50	127.00
	9	220.70	218.60	108.40	107.30	32.03	31.62	153.70	147.80
	12	275.70	281.90	111.90	112.80	35.64	33.21	159.30	158.20
	15	344.50	332.80	115.90	115.70	37.02	37.45	163.20	161.30
	18	403.20	399.70	120.40	119.10	40.69	40.25	177.10	178.70