Prokaryotic Expression and Antiserum Preparation of Functional Domain of Banana streak virus Coat Protein Gene

ACTA HORTICULTURAE SINICA ›› 2013, Vol. 40 ›› Issue (12): 2401-2408.

• Fruit Trees • Previous Articles Next Articles

Prokaryotic Expression and Antiserum Preparation of Functional Domain of Banana streak virus Coat Protein Gene

CHEN Xiu, RAO Xue-Qin, RUAN Xiao-Lei, LIU Fu-Xiu, LI Hua-Ping

（1 Boluo County Institute of Agricultural Sciences，Boluo，Guangdong 516100，China；2 Laboratory of Plant Virology，
South China Agricultural University，Guangzhou 510642，China；3 Hainan Entry-Exit Inspection and Quarantine Bureau，
Haikou 570311，China）

Online:2013-12-25 Published:2013-12-25

Abstract

Abstract: In order to provide rapid detection of Banana streak virus Guangdong isolate（BSV-GD）
and to study the virus’ gene function，BSV-GD coat protein（CP）functional domain was cloned，and
prokaryotic expression recombinant plasmid pET28b-CP was constructed by inserting the cloned gene into
the plasmid pET-28b（+）. Induced by IPTG，Escherichia coli Rosseta（DE3）containing pET28b-CP
produced the fusion protein His-CP about 46.5 kD in size. Soluble analysis of the fusion protein indicated
that it was in the inclusion body. The highly purified interest protein was obtained by using the His tag
purification kit. The special polyclonal antibody was generated through the purified protein immunizing
healthy big ear rabbits. Western-blotting analysis showed that the antiserum has very strong specificity.
Indirect enzyme immunoassay suggested the antiserum titer was higher than 1︰204 800. The available
antiserum concentration for virus detection from plant materials was 1︰1 600–1︰6 400.

Key words: banana, Banana streak virus, functional domain of coat protein gene, prokaryotic expression, polyclonal antibody

CLC Number:

S 668.1

CHEN Xiu, RAO Xue-Qin, RUAN Xiao-Lei, LIU Fu-Xiu, LI Hua-Ping. Prokaryotic Expression and Antiserum Preparation of Functional Domain of Banana streak virus Coat Protein Gene[J]. ACTA HORTICULTURAE SINICA, 2013, 40(12): 2401-2408.

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Prokaryotic Expression and Antiserum Preparation of Functional Domain of Banana streak virus Coat Protein Gene

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