Expression of the arrowhead proteinase inhibitor B (API) gene in the silkworm larvae and pupae

Ping Ji; Zhi Fang Zhang; Jia Lu He; Xiang Fu Wu; Hing Yat Lam Peter

Expression of the arrowhead proteinase inhibitor B (API) gene in the silkworm larvae and pupae

Ping Ji, Zhi Fang Zhang, Jia Lu He, Xiang Fu Wu, Hing Yat Lam Peter

Research output: Contribution to journal › Article › peer-review

Abstract

The arrowhead proteinase inhibitor B(API) gene was cloned into baculovirus transfer vectors of pBacPAK8 and pOSX, and two recombinant transfer vectors, pBacPAK (API) and pOSX(API), were constructed. Coinfection was accomplished with either one of the recombinant transfer vectors and Bm-BacPAK6 DNA in BmN cells. Then recombinant virus of CrBK9, CrBK10 and BX was selected. API gene was successfully expressed in silkworm larvae and pupae. The recombinant API(rAPI) showed strong inhibitive activity on bovine pancreas trypsin. The inhibiting activity of the expression product was 1.0 × 10 ⁵u/ml haemolymph for silkworm larvae and 1.3 × 10 ₅u/ml haemolymph for pupae. In addition, SDS-PAGE of the preliminarily purified rAPI showed its molecular weight to be about 19kD.

Original language	English (US)
Pages (from-to)	22-23
Number of pages	2
Journal	Acta Biochimica et Biophysica Sinica
Volume	29
Issue number	1
State	Published - 1997
Externally published	Yes

Keywords

Arrowhead proteinase inhibitor B gene
Gene expression
Recombinant BmNPV

ASJC Scopus subject areas

Biophysics
Biochemistry

Cite this

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title = "Expression of the arrowhead proteinase inhibitor B (API) gene in the silkworm larvae and pupae",

abstract = "The arrowhead proteinase inhibitor B(API) gene was cloned into baculovirus transfer vectors of pBacPAK8 and pOSX, and two recombinant transfer vectors, pBacPAK (API) and pOSX(API), were constructed. Coinfection was accomplished with either one of the recombinant transfer vectors and Bm-BacPAK6 DNA in BmN cells. Then recombinant virus of CrBK9, CrBK10 and BX was selected. API gene was successfully expressed in silkworm larvae and pupae. The recombinant API(rAPI) showed strong inhibitive activity on bovine pancreas trypsin. The inhibiting activity of the expression product was 1.0 × 10 5u/ml haemolymph for silkworm larvae and 1.3 × 10 5u/ml haemolymph for pupae. In addition, SDS-PAGE of the preliminarily purified rAPI showed its molecular weight to be about 19kD.",

keywords = "Arrowhead proteinase inhibitor B gene, Gene expression, Recombinant BmNPV",

author = "Ping Ji and Zhang, {Zhi Fang} and He, {Jia Lu} and Wu, {Xiang Fu} and Peter, {Hing Yat Lam}",

year = "1997",

language = "English (US)",

volume = "29",

pages = "22--23",

journal = "Acta Biochimica et Biophysica Sinica",

issn = "0582-9879",

publisher = "Shanghai Kexue Jishu Chubanshe/Shanghai Scientific & Technical Publishers",

number = "1",

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TY - JOUR

T1 - Expression of the arrowhead proteinase inhibitor B (API) gene in the silkworm larvae and pupae

AU - Ji, Ping

AU - Zhang, Zhi Fang

AU - He, Jia Lu

AU - Wu, Xiang Fu

AU - Peter, Hing Yat Lam

PY - 1997

Y1 - 1997

N2 - The arrowhead proteinase inhibitor B(API) gene was cloned into baculovirus transfer vectors of pBacPAK8 and pOSX, and two recombinant transfer vectors, pBacPAK (API) and pOSX(API), were constructed. Coinfection was accomplished with either one of the recombinant transfer vectors and Bm-BacPAK6 DNA in BmN cells. Then recombinant virus of CrBK9, CrBK10 and BX was selected. API gene was successfully expressed in silkworm larvae and pupae. The recombinant API(rAPI) showed strong inhibitive activity on bovine pancreas trypsin. The inhibiting activity of the expression product was 1.0 × 10 5u/ml haemolymph for silkworm larvae and 1.3 × 10 5u/ml haemolymph for pupae. In addition, SDS-PAGE of the preliminarily purified rAPI showed its molecular weight to be about 19kD.

AB - The arrowhead proteinase inhibitor B(API) gene was cloned into baculovirus transfer vectors of pBacPAK8 and pOSX, and two recombinant transfer vectors, pBacPAK (API) and pOSX(API), were constructed. Coinfection was accomplished with either one of the recombinant transfer vectors and Bm-BacPAK6 DNA in BmN cells. Then recombinant virus of CrBK9, CrBK10 and BX was selected. API gene was successfully expressed in silkworm larvae and pupae. The recombinant API(rAPI) showed strong inhibitive activity on bovine pancreas trypsin. The inhibiting activity of the expression product was 1.0 × 10 5u/ml haemolymph for silkworm larvae and 1.3 × 10 5u/ml haemolymph for pupae. In addition, SDS-PAGE of the preliminarily purified rAPI showed its molecular weight to be about 19kD.

KW - Arrowhead proteinase inhibitor B gene

KW - Gene expression

KW - Recombinant BmNPV

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JO - Acta Biochimica et Biophysica Sinica

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IS - 1

ER -

Expression of the arrowhead proteinase inhibitor B (API) gene in the silkworm larvae and pupae

Abstract

Keywords

ASJC Scopus subject areas

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