鸡α-2,3-唾液酸转移酶Ⅰ基因的克隆和真核表达载体的构建

Abstract

Abstract: Total RNA was extracted from intestinal of chicken by the method of Trizol. The α-2, 3-sialyltransferase Ⅰ(ST3GAL Ⅰ) gene’s cDNA was amplified by reverse transcription polymerase chain reaction with specific primer. The amplified fragment was cloned into pGEM-T easy vector, then got the recombinant plasmid. Sub-cloning the ORF of ST3GAL Ⅰ gene of pGEM-ST3GAL Ⅰ and inserted the eukaryotic expression vector pcDNA3.1(+). The target segment in recombinant plasmid was confirmed by PCR, restriction enzyme digestion and sequencing methods. The results indicated that ST3GAL Ⅰ gene was 1029 bp in length, the result of sequence was consistent with the sequence which included in GenBank, and without any mutation and deletion of codon. The length and directions of gene which inserted to eukaryotic expression vector pcDNA3.1 (+) were all correct. The recombinant eukaryotic expression vector pcDNA3.1(+)-ST3GAL Ⅰ had been constructed successfully, which laid the foundation for further eukaryotic expressing and studying of functional of ST3GAL Ⅰ gene.

Key words: avain influenza virus; receptor; α-2,3-sialyltransferase Ⅰ; eukaryotic expression vector

CLC Number:

LIU Yuan-yuan, WANG Jun-ya, ZHANG Xiao-juan, CHEN Li-peng, YUE Xu-long, GAO Wen-ming, LI Shuang-liang, CUI Bao-an, LI Xin-sheng. Cloning and Construction of Eukaryotic Expression Vector of Chicken ST3GAL Ⅰ Gene[J]. , 2013, 40(1): 21-24.

References

1 田晓滨,孙立,杨述华.克隆并构建人骨形成蛋白2真核表达载体[J].中国修复重建外科杂志,2006,2:112~115.
2 李春艳,肖晶,李曦,等.微载体规模化培养MDCK细胞增殖H9N2亚型禽流感病毒的研究[J].中国人畜共患病学报,2009,25:1149~1153.
3 库朝锋,王新华,薄新文,等. 绵羊生殖道抗菌肽cDNA克隆及真核表达载体构建[J].中国畜牧兽医, 2009, 36(6): 69~72.
4 Chu C, Lugovtsev V, Golding H, et al.Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production[J].Proc Natl Acad Sci USA,2009,106:14802~14807.
5 Connor R J, Kawaoka Y, Webster R G, et al. Receptor specificity in human,avian,and equine H2 and H3 influenza virus isolates[J].Virology,1994,295:17~23.
6 Davis G L,Hoofnggle J H. Interferon in viral hepatitis:Role in pathogenesis and treatment[J].Hepatology,1986,6:1038~1041.
7 Hatakeyama S, Sakai-Tagawa Y, Kiso M, et al. Enhanced expression of an α-2,6-linked sialic acid on MDCK cells improves isolation of human influenza viruses and evaluation of their sensitivity to a neuraminidase inhibitor[J].Clinical Microblology,2005,43:4139~4146.
8 Ito T,Suzuki Y,Takada A,et al. Differences in sialic acid-galactose linkages in the chicken egg amnion and allantois influence human influenza virus receptor specificity and variant selection[J].Virology,1997,71(4):3357~3362.
9 Ito T,Kawaoka Y.Host-range barrier of influenza A viruses[J].Vet Microbiol,2000,74:71~75.
10 Kistner O, Barrett N, Mundt W, et al. Development of a mammalian cell(Vero) derived candidate influenza virus vaccine[J]. Vaccine,1998,16:960~968.
11 Matrosovich M, Matrosovich T, Carr J, et al. Overexpression of the alpha-2,6-sialyltransferase in MDCK cells increases influenza virus sensitivity to neuraminidase inhibitors[J].Virology,2003,77:8418~8425.
12 Robertsen J S.An overview of host cell selection[J].Dev Biol Stand,1999,98:7~11.
13 Rogers G N, Paulson J C. Receptor determinants of human and animal influenza virus isolate:Differences in receptor specificity of the H3 hemagglutinin based on species of origin[J].Virology,1983,127:361~373.
14 Sun X, Tan W, Zhang Y, et al. Effect of the seeding density on the growth of Vero cells on microcarriers[J].East China University of Science and Technology,1999,25:367~370.
15 Trebbien R, Larsen L S, Viuff B M. Distribution of sialic acid receptors and influenza A virus of avian and swine origin in experimentally infected pigs[J].Virology,2011,8:434~448.

[1]	PAN Pengcheng, LEI Zongquan, LI Xian, HU Xiangyun, QIN Qiantao, QIN Zhaoxian, GUAN Zhihui, CHEN Baojian, XIE Bingkun. Bioinformatics Analysis,Eukaryotic Expression Vector Construction and Tissue Expression of MYL2 Gene in Luchuan Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1796-1806.
[2]	ZHANG Guoqing, JIANG Yuhang, ZHANG Shuang, LI Letian, HAO Jiayi, CHEN Jing, ZOU Wancheng, LI Chang, LI Taiyuan, GAO Xu, JIN Ningyi. Preparation and Identification of PDCoV RBD Protein Peptide Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1575-1583.
[3]	LIANG Jingwen, WANG Lina. Research Progress of Bile Acid Function in Gut-brain Axis [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 924-931.
[4]	LEI Qi, REN Jie, KANG Jie. Effects of Scutellaria baicalensis georgi on Antioxidant Capacity,Sex Hormone Levels and mRNA Expression of ERα in Female Mice Exposed to ZEA [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 989-997.
[5]	XIE Jingjing, REN Mengting, FAN Jiahui, JIAO Yaru, HAN Zongxi, MA Deying. Study on Immune Response of Infectious Bronchitis Virus to Chickens and Antiviral Effect of Recombinant Avian Beta-defensins 11 and 12 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 246-259.
[6]	ZHOU Ya, ZHANG Heyang, LIU Linling, LI Haodong, ZHENG Junjun, WANG Guiwu. Analysis of FGFR2 Gene Polymorphism and Their Correlation with Antler Weight Traits in Sika Deer [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3006-3014.
[7]	YU Kai, ZHAO Yufen, WENG Yu, WANG Qin, HAO Shaoyu, YU Boyang, DU Chenguang, SUBUDENG Gerile, LI Haijun. Effects of Dihydrotestosterone on Proliferation and Expression of Anti-Müllerian Hormone of Mouse Granulosa Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3036-3043.
[8]	LI Weidong, JIANG Yuting, DUAN Hongwei, HE Haijun, YANG Shuai, DING Ziqiang, WU Jianxin, ZHANG Enqiu, HU Junjie. Distribution and Expression of Melatonin Receptors MT1 and MT2 in Various Stomach Tissues of Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 3256-3262.
[9]	CHEN Shiyu, CUI Shuai, JIANG Yajun, XIN Ting, WANG Yang, HAO Yuxin, HUANG Jianxin, GUO Xiaoyu, ZHU Hongfei, WU Jiajun, JIA Hong. lncRNA-mRNA Analysis of Toll-like Receptor Signaling in Porcine Peripheral Blood Lymphocytes After African Swine Fever Virus Infection [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2677-2687.
[10]	ZOU Jianwei, ZOU Juhong, SHEN Yujian, WEI Yi, ZHANG Sanbao, LIAN Zitong, XU Jianjian, SONG Ying, HUANG Yanna, WEI Yingming, JIANG Qinyang, ZHENG Zihua. Bioinformatics Analysis, Construction of Eukaryotic Expression Vector and Expression of LMCD1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2033-2042.
[11]	LI Peiqi, KANG Jie, WANG Keyao, WANG Huimin, HAN Yufeng, WANG Tinghui, CUI Liang, ZHANG Jian, DENG Ruiqiang, DUAN Zhibian. Effect of Compound Traditional Chinese Medicine Water Extract on TGF-β1/Smad Pathway in Liver of Ascites Syndrom in Broilers [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1860-1869.
[12]	SHI Qian, XU Xinfeng, LI Xuewen, XIAO Yuanyuan, BAI Meihua, ZHANG Fuchun. Research Progress on the Mechanism of Polysaccharide as Adjuvant of Veterinary Vaccine [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1895-1907.
[13]	CUI Chengyu, MA Ben, ZHAO Jiahui, ZHAO Zijuan, CHEN Yu, CAO Zhongzan, LUAN Xinhong. Application Prospect of Adiponectin and Its Receptor Agonists in Avian Fatty Liver Disease [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 738-745.
[14]	WU Shuangmin, LI Long, HOU Ren, CHEN Yushuang, PENG Dapeng. Research Progress on the Interaction Between Small Molecules of Veterinary Drugs and Biometric Elements [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(2): 755-764.
[15]	SHAO Shuyu, LI Jia, MA Zhuo, YU Chang, ZHANG Ying, ZHANG Jinlong, MA Zhiyu. Construction and Identification of Lentiviral Overexpression Vector of Porcine Neuromedin B Receptor Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(10): 3756-3763.

Cloning and Construction of Eukaryotic Expression Vector of Chicken ST3GAL Ⅰ Gene

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments