The allantoic fluid was collected from the dead chicken embryo within 48-96 hours after the 11-day-old SPF brambling chicken embryos by type ZH-1. To amplify the sequence of the fusion gene( F- gene) and to determine whether the amplified sequence contains the integrated sequence of open reading frame(ORF),one pair of specific primer was designed and synthesized basedon the published sequence of the F gene and then was used to amplify the F-gene in brambling by RT-PCR. The amplified fragment was taken back and purified, and then was cleaved by both EcoR I and Sal I enzymes. Later,the F gene was transformed into DH5αby PCI-neo vector. The detected results of positive isolates by both PCRanalysis and enzyme cleaving response with EcoRI and Sal I,which showed that the expression vector PCI- neo-F has been constructed successfully,and thispaved the way to study the Vero cell of mammals. The information about the amplified sequence and the compared knowledge in nucleotide composition, amino acid sequence and systemically de- velopment tree between the amplified F gene sequence and the other ZH-1 F gene,were useful for the further elucidate the genetic background of paramyxovirus type ZH-1.
LI Hong li
,
ZHAN Li e
,
QIAo Zhong
,
WANG Cai xian
,
LU Bing yang
. Construction of the F Gene Eukaryotic Expression Vector of the Paramyxovirus Strain ZH-1 From Brambling[J]. Acta Agriculturae Boreali-Sinica, 2009
, 24(3)
: 54
-58
.
DOI: 10.7668/hbnxb.2009.03.012
[1] 梅双双,杨润德.鸡新城疫病毒强毒株的分离及生物学特性鉴定[J].中国预防兽医学报,2002,5(1):368-370.
[2] 詹丽娥,乔忠,薛俊龙.H9N2亚型禽流感病毒株的分离和鉴定[J].山西农业科学,2004,32(4):74-76.
[3] 殷震,刘景华.动物病毒学[M].北京:科学出版社,1997:326-436.
[4] Chen J P,Wang C H.Phylagenetie analysis of Newcastle dis-ease virus in Taiwan[J].Jmicrobiol Immun Infect,2002,35(4):223-228.
[5] 毕玉海,曹璐,李志杰,等.鹅副粘病毒与鹅细小病毒主要免疫原性基因在bae-to-bac杆状病毒表达系统中的共表达及其免疫[J].中国兽医学报,2008,28(2):125-127.
[6] 费恩阁,李德昌,丁壮.动物物疫病学[M].北京:中国农业出版社,2004:506-513.
[7] 王学理,武迎红,丁壮.鹅源禽副牯病毒NA-1株生物学特性的研究[J].吉林农业大学学报,2006,28(1):73-97.
[8] 丁壮,金宁一,王兴龙,等.新城疫病毒四平株HN蛋白基因的克隆与序列分析[J].中国预防兽医学报,2000.22(2):102-105.
[9] 王学理,丁壮,左玉柱,等.鹅副粘病毒NA-1分离株HN蛋白基因的克隆与序列分析[J].中国预防兽医学报,2005,27(1):18-21.
[10] 刘华雷,王永坤,朱国强,等.一株基因Ⅶ型新城疫病毒的分离及分子鉴定[J].中国兽医学报,2002,22(3):216-218.
[11] Huang Z,Panda h,Elankumaran S,et al.The hemagglu-tinin-neuraminidase protein of newcastle disease virus deter-mines tropism and virulence[J].J Virol,2004,78(8):4176-4184.
[12] 符芳,姜北宇,张莉,等.新城疫病毒La._qota F基因克隆及原核表达[J].华北农学报,2006,21(3):117-120.
[13] 毕玉海,丛彦龙,李志杰,等.猪源副粘病毒的分子鉴定[J].中国兽医学报,2008,28(4):343-349.
[14] 王会中,付秋霞,高明,等.小鼠脂联素与绿色荧光蛋白融合基因表达载体的构建及在COS-7细胞中的表达[J].生物技术通讯,2005,16(5):492-495.
[15] 由振强,于涟,翁继锋,等.鸡白细胞介素2真核表达质粒的构建、表达及对AIV疫苗免疫增强作用研究[J].浙江农业学报,2005,17(6):350-353.
[16] 王福祥,孙永涛,孙永年,等.中国株HIV.1 gag基因核酸疫苗免疫小鼠的实验研究[J].解放军医学杂志.2004,29(5):430-432.
[17] Hulse D J,Romero c H.Partial protection against infectious bursal disease virns through DNA mediated vaccination with theVP2 capsid Drotein and chicken IL-2 genes[J].Vac.cine.2004.22(9-10):1249-1259.
