Prokaryotic Expression of Chicken Angiopoietin-Like 4 Recombinant Protein and Its Effects on Serum Biochemical Indices and Hormone Levels of Broilers

ZHAO Xu; HUANG Huashan; SU Shanshan

doi:10.3969/j.issn.1006-267x.2020.04.045

2020 , Vol. 32 >Issue 4: 1859 - 1866

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.04.045

Prokaryotic Expression of Chicken Angiopoietin-Like 4 Recombinant Protein and Its Effects on Serum Biochemical Indices and Hormone Levels of Broilers

ZHAO Xu ,
HUANG Huashan ,
SU Shanshan

Expand

1. College of Agriculture and Forestry Science, Linyi University, Linyi 276000, China;
2. Shandong Longda Biotechnology Co., Ltd., Linyi 276400, China;
3. Shanghai Novoprotein Scientific Co., Ltd., Shanghai 201203, China

Received date: 2019-09-26

Online published: 2020-04-16

Supported by

Fold

Abstract

The aim of this study was to accomplish the prokaryotic expression of chicken angiopoietin-like protein 4 (ANGPTL4) recombinant protein, and to investigate the effects of this recombinant protein on serum biochemical indices and hormone levels of broilers. The gene of chicken ANGPTL4 with a His-SUMO tag was cloned into Nde Ⅰ and Xho Ⅰsites of pET21a(+) vector to construct recombinant plasmid pET21a-His-SUMO-ANGPTL4. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and the bacteria were induced with isopropy-β-D-thiogalactoside (IPTG). After inclusion body refolding, the refold protein was purified by Ni column and the expected size of fusion protein His-SUMO-ANGPTL4 was obtained. Thirty-six broilers at 35 days of age were subjected to one of the six following treatments:intravenous injection of chicken ANGPTL4 recombinant protein at the doses of 0 (control), 20, 100, 500, 2 500 and 12 500 ng/kg BW. Six broilers for each dose, and fasting blood samples of broilers were collected at 30 min after injection. The results showed as follows:1) compared with the control group, the contents of glucose (GLU) and very low-density lipoprotein (VLDL) in the serum of 2 500 and 12 500 ng/kg BW chicken ANGPTL4 recombinant protein groups were significantly increased (P<0.05). As the injected dose of chicken ANGPTL4 recombinant protein increased from 0 to 12 500 ng/kg BW, the contents of GLU and VLDL in the serum of broilers were linearly (P<0.05) and quadratically (P<0.05) increased. 2) Compared with the control group, the growth hormone (GH) level in the serum of 20, 2 500 and 12 500 ng/kg BW chicken ANGPTL4 recombinant protein groups were significantly decreased (P<0.05). As the injected dose of chicken ANGPTL4 recombinant protein increased from 0 to 12 500 ng/kg BW, the GH level in the serum of broilers were linearly (P<0.05) and quadratically (P<0.05) decreased. 3) Compared with the control group, the leptin level in the serum of 20, 100, 500, 2 500 and 12 500 ng/kg BW chicken ANGPTL4 recombinant protein groups were significantly decreased (P<0.05). It is concluded that the prokaryotic expression of chicken ANGPTL4 recombinant protein is successfully constructed in this study, and intravenous injection of this recombinant protein at the doses of 2 500 and 12 500 ng/kg BW can significantly increase the serum GLU and VLDL contents, and significantly decrease the serum GH and leptin levels of broilers.

Key words： chicken ANGPTL4 recombinant protein; prokaryotic expression; broilers; serum biochemical indices; serum hormone levels

Cite this article

ZHAO Xu , HUANG Huashan , SU Shanshan . Prokaryotic Expression of Chicken Angiopoietin-Like 4 Recombinant Protein and Its Effects on Serum Biochemical Indices and Hormone Levels of Broilers[J]. Chinese Journal of Animal Nutrition, 2020 , 32(4) : 1859 -1866 . DOI: 10.3969/j.issn.1006-267x.2020.04.045

References

[1] MANDARD S,ZANDBERGEN F,TAN N S,et al.The direct peroxisome proliferator-activated receptor target fasting-induced adipose factor (FIAF/PGAR/ANGPTL4) is present in blood plasma as a truncated protein that is increased by fenofibrate treatment[J].The Journal of Biological Chemistry,2004,279(33):34411-34420.

[2] GROOTAERT C,VAN DE WIELE T,VERSTRAETE W,et al.Angiopoietin-like protein 4:health effects,modulating agents and structure-function relationships[J].Expert Review of Proteomics,2012,9(2):181-199.

[3] ZHAO X,GUO Y M,LIU H B,et al.Clostridium butyricum reduce lipogenesis through bacterial wall components and butyrate[J].Applied Microbiology and Biotechnology,2014,98(17):7549-7557.

[4] ZHAO X,DING X,YANG Z B,et al.Effects of Clostridium butyricum on breast muscle lipid metabolism of broilers[J].Italian Journal of Animal Science,2018,17(4):1010-1020,doi:10.1080/1828051X.2018.1453758.
[5] 王镜岩,朱圣庚,徐长法.生物化学(上册)[M].3版.北京:高等教育出版社,2002:118-119.
[6] BÄCKHED F,DING H,WANG T,et al.The gut microbiota as an environmental factor that regulates fat storage[J].Proceedings of the National Academy of Sciences of the United States of America,2004,101(44):15718-15723.

[7] GOLDBERG I J.Lipoprotein lipase and lipolysis:central roles in lipoprotein metabolism and atherogenesis[J].Journal of Lipid Research,1996,37(4):693-707.
[8] 赵旭.丁酸梭菌对肉鸡脂肪代谢的影响及其机理研究[D].博士学位论文.北京:中国农业大学,2014:16-17.
[9] GE H F,YANG G Q,YU X X,et al.Oligomerization state-dependent hyperlipidemic effect of angiopoietin-like protein 4[J].Journal of Lipid Research,2004,45(11):2071-2079.

[10] KÖSTER A,CHAO Y B,MOSIOR M,et al.Transgenic angiopoietin-like (ANGPTL)4 overexpression and targeted disruption of ANGPTl4 and ANGPTl3:regulation of triglyceride metabolism[J].Endocrinology,2005,146(11):4943-4950.

[11] MANDARD S,ZANDBERGEN F,VAN STRATEN E,et al.The fasting-induced adipose factor/angiopoietin-like protein 4 is physically associated with lipoproteins and governs plasma lipid levels and adiposity[J].Journal of Biological Chemistry,2006,281(2):934-944.

[12] LICHTENSTEIN L,BERBÉE J F P,VAN DIJK S J,et al.ANGPTl4 upregulates cholesterol synthesis in liver via inhibition of LPL-and HL-dependent hepatic cholesterol uptake[J].Arteriosclerosis,Thrombosis,and Vascular Biology,2007,27(11):2420-2427.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References