猪唾液酸黏附素氨基端结构域基因的克隆表达及其多克隆抗体制备

Abstract

Abstract: The pig sialoadhesin near the amino structure domain cDNA sequence which was cloned from the 90-day age lung health piglets macrophages with RT-PCR was subcloned into a prokaryotic expression vector pET-32a, to construct a recombinant expression plasmid pET-Sn150 which was successfully used to express a gamma recombinant protein subunits whose molecular weight was about 43 ku. By using the recombinant protein pET-Sn150 to immune the mouse, the mice sialoadhesin resistance gamma-irradiation His recombinant protein polyclonal antibody which could be tested by Western blotting and indirect ELISA could be achieved. And the result of indirect ELISA showed that the titer of the polyclonal antibody was 1∶12800. The specific binding of the mice sialoadhesin resistance gamma-irradiation His recombinant protein polyclonal antibody with the restructuring γ and unit protein revealed by the result of Western blotting proved that the recombinant protein had good immunogenicity, and that layed the foundation for a further study of its construction and function as the pig sialoadhesin near the amino structure domain gene was successfully cloned and expressed.

Key words: sialoadhesin; gene clone; prokaryotic expression; polyclonal antibody 

CLC Number:

Q786

ZHANG Ji-xi, ZHANG Zhi-yuan, ZHANG Yi-na, ZHOU Yong-hui, ZHANG Xiang, HAO Yi-mei, ZHANG Yuan-yuan, XIA Ping-an, CUI Bao-an. Cloning and Expression of the Amino Structure Domain Gene of Sialoadhesin from Swine and Preparation of Polyclonal Antibody against it[J]. .

References

1 Abin M F, Spronk G, Wagner M. Comparative infection efficiency of porcine reproductive and respiratory syndrome virus field isolates on MA104 cells and porcine alveolar macrophages[J]. Can J Vet Res, 2009,73(3):4~200.

2 Cano J P, Dee S A, Murtaugh M P. Infection dynamics and clinical manifestations following experimental inoculation of gilts at 90 days of gestation with a low dose of porcine reproductive and respiratory syndrome virus[J].Can J Vet Res, 2009,73(4):38~79.

3 Dee S, Otake S, Oliveira S. Evidence of long distance airborne transport of porcine reproductive and respiratory syndrome virus and mycoplasma hyopneumoniae[J]. Vet Res, 2009,40(4):39~89.

4 Delrue I, Delputte P L, Nauwynck H J. Assessing the functionality of viral entry-associated domains of porcine reproductive and respiratory syndrome virus during inactivation procedures, a potential tool to optimize inactivated vaccines[J]. Vet Res, 2009,40(6):62~126.

5 Kinch M S, Yunus A S, Lear C. A broad-spectrum small molecule inhibitor of viral infection[J]. Am J Transl Res, 2009, 4(1):87~98.

6 Klinge K L, Vaughn E M. Age-dependent resistance to porcine reproductive and respiratory syndrome virus replication in swine[J]. Virol J, 2009, 27 (1):67~88.

7 Nedialkova D D, Ulferts R, Lauber C. Biochemical characterization of arterivirus nonstructural protein 11 reveals the nidovirus-wide conservation of a replicative endoribonuclease[J]. J Virol, 2009,83(11):56~82.

8 Pitkin A, Deen J, Otake S. Further assessment of houseflies(Musca domestica) as vectors for the mechanical transport and transmission of porcine reproductive and respiratory syndrome virus under field conditions[J]. Can Vet Res, 2009,73(2):23~69.

9 Rosenfeld P, Turner P V, Macinnes J I. Evaluation of porcine reproductive and respiratory syndrome virus replication in laboratory rodents[J].Can J Vet Res, 2009,73(4):30~90.

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Cloning and Expression of the Amino Structure Domain Gene of Sialoadhesin from Swine and Preparation of Polyclonal Antibody against it

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