Molecular cloning of a female-specific cDNA with unique repeat sequences from the fat body of the adult locust, Locusta migratoria

Qili Feng; Subba R. Palli; Tim R. Ladd; Sardar S. Sohi; Arthur Retnakaran; Kenneth G. Davey

doi:10.1016/S0965-1748(00)00055-2

Molecular cloning of a female-specific cDNA with unique repeat sequences from the fat body of the adult locust, Locusta migratoria

Qili Feng, Subba R. Palli, Tim R. Ladd, Sardar S. Sohi, Arthur Retnakaran, Kenneth G. Davey

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

1 Scopus citations

Abstract

A cDNA clone encoding a 25-kDa protein (25K) was isolated from a cDNA library made from RNA isolated from the adult fat body and ovaries of the locust, Locusta migratoria. The longest open reading frame of this cDNA clone encodes a 225-amino acid polypeptide, the N-terminal end of which was similar to the 21-kDa and 19-kDa juvenile hormone induced proteins identified in the locust hemolymph, but the C-terminal end was different. The C-terminal end of the 25K cDNA contained seven unique repeat elements of 10 amino acids each, most of which are polar residues. Expression of the 25K mRNA was tissue-, development- and sex-specific. A 1.2-kb mRNA was detected using the 25K cDNA as a probe only in the fat body of adult females. The mRNA started to appear at day 4 after the insect molted to the adult and rapidly increased by day 6. The mRNA was absent in the ovarian follicle cells and fat body of adult males. In vitro transcription and translation of the 25K cDNA produced a protein that migrated around 32 kDa on sodium dodecyl sulfate polyacrylamide gels. The 25K cDNA was expressed in a baculovirus expression system and the protein produced also migrated around 32 kDa. (C) 2000 Elsevier Science Ltd.

Original language	English
Pages (from-to)	829-837
Number of pages	9
Journal	Insect Biochemistry and Molecular Biology
Volume	30
Issue number	8-9
DOIs	https://doi.org/10.1016/S0965-1748(00)00055-2
State	Published - Sep 2000

Bibliographical note

Funding Information:
This research was supported in part by a Natural Sciences and Engineering Research Council of Canada Research Grant, and a Collaborative Research and Development Grant from NSERC and Cyanamid Canada to K.G.D. and by the National Biotechnology Strategy Fund and the Science and Technology Opportunities Fund of the Canadian Forest Service to Biotechnology at GLFC.

Funding

This research was supported in part by a Natural Sciences and Engineering Research Council of Canada Research Grant, and a Collaborative Research and Development Grant from NSERC and Cyanamid Canada to K.G.D. and by the National Biotechnology Strategy Fund and the Science and Technology Opportunities Fund of the Canadian Forest Service to Biotechnology at GLFC.

Funders	Funder number
Canadian Forest Service to Biotechnology
Cyanamid Canada
National Biotechnology Strategy Fund
Natural Sciences and Engineering Research Council of Canada

Keywords

Baculovirus expression system
Fat body
Follicle cell
Hemolymph
Repeat element
Reproductive maturation

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Insect Science

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10.1016/S0965-1748(00)00055-2

Cite this

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title = "Molecular cloning of a female-specific cDNA with unique repeat sequences from the fat body of the adult locust, Locusta migratoria",

abstract = "A cDNA clone encoding a 25-kDa protein (25K) was isolated from a cDNA library made from RNA isolated from the adult fat body and ovaries of the locust, Locusta migratoria. The longest open reading frame of this cDNA clone encodes a 225-amino acid polypeptide, the N-terminal end of which was similar to the 21-kDa and 19-kDa juvenile hormone induced proteins identified in the locust hemolymph, but the C-terminal end was different. The C-terminal end of the 25K cDNA contained seven unique repeat elements of 10 amino acids each, most of which are polar residues. Expression of the 25K mRNA was tissue-, development- and sex-specific. A 1.2-kb mRNA was detected using the 25K cDNA as a probe only in the fat body of adult females. The mRNA started to appear at day 4 after the insect molted to the adult and rapidly increased by day 6. The mRNA was absent in the ovarian follicle cells and fat body of adult males. In vitro transcription and translation of the 25K cDNA produced a protein that migrated around 32 kDa on sodium dodecyl sulfate polyacrylamide gels. The 25K cDNA was expressed in a baculovirus expression system and the protein produced also migrated around 32 kDa. (C) 2000 Elsevier Science Ltd.",

keywords = "Baculovirus expression system, Fat body, Follicle cell, Hemolymph, Repeat element, Reproductive maturation",

author = "Qili Feng and Palli, {Subba R.} and Ladd, {Tim R.} and Sohi, {Sardar S.} and Arthur Retnakaran and Davey, {Kenneth G.}",

note = "Funding Information: This research was supported in part by a Natural Sciences and Engineering Research Council of Canada Research Grant, and a Collaborative Research and Development Grant from NSERC and Cyanamid Canada to K.G.D. and by the National Biotechnology Strategy Fund and the Science and Technology Opportunities Fund of the Canadian Forest Service to Biotechnology at GLFC.",

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AU - Feng, Qili

AU - Palli, Subba R.

AU - Ladd, Tim R.

AU - Sohi, Sardar S.

AU - Retnakaran, Arthur

AU - Davey, Kenneth G.

N1 - Funding Information: This research was supported in part by a Natural Sciences and Engineering Research Council of Canada Research Grant, and a Collaborative Research and Development Grant from NSERC and Cyanamid Canada to K.G.D. and by the National Biotechnology Strategy Fund and the Science and Technology Opportunities Fund of the Canadian Forest Service to Biotechnology at GLFC.

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AB - A cDNA clone encoding a 25-kDa protein (25K) was isolated from a cDNA library made from RNA isolated from the adult fat body and ovaries of the locust, Locusta migratoria. The longest open reading frame of this cDNA clone encodes a 225-amino acid polypeptide, the N-terminal end of which was similar to the 21-kDa and 19-kDa juvenile hormone induced proteins identified in the locust hemolymph, but the C-terminal end was different. The C-terminal end of the 25K cDNA contained seven unique repeat elements of 10 amino acids each, most of which are polar residues. Expression of the 25K mRNA was tissue-, development- and sex-specific. A 1.2-kb mRNA was detected using the 25K cDNA as a probe only in the fat body of adult females. The mRNA started to appear at day 4 after the insect molted to the adult and rapidly increased by day 6. The mRNA was absent in the ovarian follicle cells and fat body of adult males. In vitro transcription and translation of the 25K cDNA produced a protein that migrated around 32 kDa on sodium dodecyl sulfate polyacrylamide gels. The 25K cDNA was expressed in a baculovirus expression system and the protein produced also migrated around 32 kDa. (C) 2000 Elsevier Science Ltd.

KW - Baculovirus expression system

KW - Fat body

KW - Follicle cell

KW - Hemolymph

KW - Repeat element

KW - Reproductive maturation

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ER -

Molecular cloning of a female-specific cDNA with unique repeat sequences from the fat body of the adult locust, Locusta migratoria

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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