基因Ⅶ型新城疫病毒F蛋白在昆虫细胞中的表达及免疫效力研究

doi:10.16431/j.cnki.1671-7236.2017.06.037

Abstract

Abstract:

This experiment was conducted to study the protein expression of F gene of Newcastle disease virus (NDV) type Ⅶ by baculovirus expression system.The F gene was amplified by RT-PCR and cloned into pFastBac HT A plasmid, and then the recombinant plasmid was transformed into DH10Bac competent cells to get the positive recombinant bacmid.After 72 h transfection of Sf9 cell with recombinant bacmid, the expression of interest protein was detected by indirect immunofluorescence assay (IFA), SDS-PAGE analysis and Western blotting. Serum antibody titers of immunized SPF chickens were determined by ELISA and the protective properties were determined by protection test. The results showed that F gene was expressed in Sf9 cells infected with the recombinant baculovirus. The expressed protein could induce high titer specific antibody against NDV. The protective rate of recombinant F protein group was 90%, which was significantly higher than that of the negative control group. These results laid a foundation for study on NDV subunit vaccine.

Key words: Newcastle disease virus; fusion protein F; recombinant baculovirus; insect cell; immune effect

CLC Number:

S855.3

SUN Peng. Study on Expression and Immune Effect of F Protein of Newcastle Disease Virus Type Ⅶ in Insect Cell[J]. , 2017, 44(6): 1840-1846.

References

[1] 甘孟侯.中国禽病学[M].北京:中国农业出版社,1997.
[2] 胡思科,席瑞珍,任涛,等.4株新城疫病毒F蛋白和HN蛋白的计算机模建与表面抗原分析[J].华南农业大学学报,2013,34(1):87-92.
[3] Saif Y M.苏敬良,高福,索勋,译.禽病学[M].第11版.北京:中国农业出版社,2005.
[4] Lomniczi B, Wehmann E, Herczeg J,et a1.Neweastle disease outbreaks in recent years in Western Europe were caused by an old (V1) and a novel genotype(Ⅶ)[J].Arch Virol,1998,143(1):49-64.
[5] 王永坤,严维巍,周继宏,等.鸡副粘病毒病病毒特性和基因型的研究及防制[J].动物科学与动物医学,2000,17(3):31-34.
[6] 石星明,王玫,张晶,等.CpG基序对新城疫病毒DNA免疫效果的影响[J].中国预防兽医学报,2010,32(7):554-558.
[7] 冯淑萍,石伟革,孙翔翔,等.不同动物来源的新城疫病毒F基因序列分析[J].动物医学进展,2014,35(12):36-40.
[8] 徐钰,解林红,耿海东,等.两株野鸭源新城疫病毒分离株F基因和HN基因的分子特性分析[J].动物医学进展,2014,35(8):65-69.
[9] Futatsumori-Sugai M, Tsumoto K. Signal peptide for improving recombinant protein secretion in the baculovirus expression vector system[J].Biochem,2010,391(1):931-935.
[10] Kathy H, Someet N, Sowmya P, et al. False positive reactivity of recombinant,diagnostic, glycoproteins produced in high five insect cell:Effect of glycosylation[J].Immunol Methods,2008,330(1-2):130-136.
[11] 王桂军,秦爱建,邵红霞,等.新城疫病毒F蛋白在昆虫细胞中的表达及其融细胞作用[J].中国病毒学,2006,21(6):571-574.
[12] 贾珊珊,孙凌霜,齐岩,等. H7亚型禽流感病毒HA1基因在重组杆状病毒中的表达及其表达产物的抗原性[J].中国兽科学, 2007, 37(3):232-235.
[13] 刘开春,孟春春,陈素娟,等.两株不同宿主来源的基因Ⅶ型新城疫病毒的生物学特性比较及全基因组序列分析[J].中国预防兽医学报,2015,37(2):89-92.
[14] 孙敏华,董嘉文,李林林,等.一株基因Ⅶ b亚型新城疫病毒F和HN基因的序列分析[J].中国预防兽医报,2013,35(1):74-76.
[15] 刘佳佳,严专强,刘迪,等.新城疫病毒广东基因Ⅶ型分离株的分离鉴定及F基因克隆与序列分析[J].动物医学进展,2013,34(10):50-53.
[16] 金宁一,鲁会军,金扩世.新城疫病毒变异进化与跨种感染[J].中国家禽,2010,32(7):1-5.
[17] 查云峰,徐兴然,肖昌,等.猪瘟病毒E2基因在昆虫细胞中的分泌表达及活性检测[J].畜牧兽医学报,2005, 36(10):1100-1105.
[18] 王善辉,谢金文,王文秀,等. NDV江苏分离株F基因的克隆与序列分析[J].中国动物检疫,2014,31(3):64-67.
[19] 游猛,潘志明,耿士忠,等.新城疫病毒基因在大肠杆菌中的高效表达及其免疫原性分析[J].中国家禽,2009, 31(20):21-24.
[20] 王兴龙,金宁一,丁壮,等.新城疫病毒F基因在新型杆状病毒表达系统中的表达及重组病毒的免疫原性[J].中国兽医学报,2001,21(6):533-535.

Study on Expression and Immune Effect of F Protein of Newcastle Disease Virus Type Ⅶ in Insect Cell

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