A λgt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin

Andres G. Grandea; Le Kim Tuyen; Nur Asikin; Theodore B. Davis; Mario Philipp; Carolyn Cohen; Larry A. McReynolds

doi:10.1016/0166-6851(89)90139-4

A λgt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin

Andres G. Grandea, Le Kim Tuyen, Nur Asikin, Theodore B. Davis, Mario Philipp, Carolyn Cohen, Larry A. McReynolds

Department of Biochemistry and Molecular Biology Pre Clinic

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

28 Citations (Scopus)

Abstract

The cDNA synthesized from mRNA of Dirofilaria immitis female adult worms was cloned into the expression vector λgt11. Screening the library with a hyperimmune rabbit antiserum raised against adult worm homogenates yielded several antigen positive clones. One of these clones, λcDi2, was recognized by rabbit antisera raised against either D. immitis L-3, adult, Brugia malayi L-3 or Onchocerca volvulus adult worm antigen, as well as by antisera from humans naturally infected with O. volvulus or Wuchereria bancrofti. Affinity-purified anti-λcDi2 antibodies reacted with a 97-kDa protein on Western transfers of adult D. immitis antigen extracts that were reduced with β-mercaptoethanol. The whole rabbit anti-D. immitis adult antiserum depleted of anti-λcDi2 antibodies exhibited decreased reactivity to this 97-kDa band. A monoclonal antibody (IA6) that specifically binds Schistosoma mansoni paramyosin also recognised a 97-kDa protein in D. immitis extracts upon Western transfer. The deduced amino acid sequence of partial DNA sequence from λcDi2 showed some similarity to nematode myosin, and gave a stretch of 82 amino acids that is 91.5% identical to Caenorhabditis elegans paramyosin: thus, λcDi2 encodes D. immitis paramyosin.

Original language	English
Pages (from-to)	31-41
Number of pages	11
Journal	Molecular and Biochemical Parasitology
Volume	35
Issue number	1
DOIs	https://doi.org/10.1016/0166-6851(89)90139-4
Publication status	Published - 1 Jun 1989

Keywords

Affinity-purified antibody
Dirofilaria immitis
Myosin
Paramyosin
λgt11 cDNA library

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10.1016/0166-6851(89)90139-4

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title = "A λgt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin",

abstract = "The cDNA synthesized from mRNA of Dirofilaria immitis female adult worms was cloned into the expression vector λgt11. Screening the library with a hyperimmune rabbit antiserum raised against adult worm homogenates yielded several antigen positive clones. One of these clones, λcDi2, was recognized by rabbit antisera raised against either D. immitis L-3, adult, Brugia malayi L-3 or Onchocerca volvulus adult worm antigen, as well as by antisera from humans naturally infected with O. volvulus or Wuchereria bancrofti. Affinity-purified anti-λcDi2 antibodies reacted with a 97-kDa protein on Western transfers of adult D. immitis antigen extracts that were reduced with β-mercaptoethanol. The whole rabbit anti-D. immitis adult antiserum depleted of anti-λcDi2 antibodies exhibited decreased reactivity to this 97-kDa band. A monoclonal antibody (IA6) that specifically binds Schistosoma mansoni paramyosin also recognised a 97-kDa protein in D. immitis extracts upon Western transfer. The deduced amino acid sequence of partial DNA sequence from λcDi2 showed some similarity to nematode myosin, and gave a stretch of 82 amino acids that is 91.5% identical to Caenorhabditis elegans paramyosin: thus, λcDi2 encodes D. immitis paramyosin.",

keywords = "Affinity-purified antibody, Dirofilaria immitis, Myosin, Paramyosin, λgt11 cDNA library",

author = "Grandea, {Andres G.} and Tuyen, {Le Kim} and Nur Asikin and Davis, {Theodore B.} and Mario Philipp and Carolyn Cohen and McReynolds, {Larry A.}",

note = "Funding Information: The authors thank Dr. D. Comb for encouragement, enthusiasm, and support, Drs. E. Pearce, A. Sher, and T. Nutman for antisera and monoclonal antibodies, Ms. M.-P. LeGuin for excellent assistance on sequence analysis, and Drs. P. Arasu, R. Limberger, C. Maina, and F. Perler for commentary on the manuscript. Dr. Cohen's lab is supported by NIH grant AM17346, NSF grant DMB85-00233, and a grant from the Muscular Dystrophy Foundation.",

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doi = "10.1016/0166-6851(89)90139-4",

language = "English",

volume = "35",

pages = "31--41",

journal = "Molecular and Biochemical Parasitology",

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T1 - A λgt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin

AU - Grandea, Andres G.

AU - Tuyen, Le Kim

AU - Asikin, Nur

AU - Davis, Theodore B.

AU - Philipp, Mario

AU - Cohen, Carolyn

AU - McReynolds, Larry A.

N1 - Funding Information: The authors thank Dr. D. Comb for encouragement, enthusiasm, and support, Drs. E. Pearce, A. Sher, and T. Nutman for antisera and monoclonal antibodies, Ms. M.-P. LeGuin for excellent assistance on sequence analysis, and Drs. P. Arasu, R. Limberger, C. Maina, and F. Perler for commentary on the manuscript. Dr. Cohen's lab is supported by NIH grant AM17346, NSF grant DMB85-00233, and a grant from the Muscular Dystrophy Foundation.

PY - 1989/6/1

Y1 - 1989/6/1

N2 - The cDNA synthesized from mRNA of Dirofilaria immitis female adult worms was cloned into the expression vector λgt11. Screening the library with a hyperimmune rabbit antiserum raised against adult worm homogenates yielded several antigen positive clones. One of these clones, λcDi2, was recognized by rabbit antisera raised against either D. immitis L-3, adult, Brugia malayi L-3 or Onchocerca volvulus adult worm antigen, as well as by antisera from humans naturally infected with O. volvulus or Wuchereria bancrofti. Affinity-purified anti-λcDi2 antibodies reacted with a 97-kDa protein on Western transfers of adult D. immitis antigen extracts that were reduced with β-mercaptoethanol. The whole rabbit anti-D. immitis adult antiserum depleted of anti-λcDi2 antibodies exhibited decreased reactivity to this 97-kDa band. A monoclonal antibody (IA6) that specifically binds Schistosoma mansoni paramyosin also recognised a 97-kDa protein in D. immitis extracts upon Western transfer. The deduced amino acid sequence of partial DNA sequence from λcDi2 showed some similarity to nematode myosin, and gave a stretch of 82 amino acids that is 91.5% identical to Caenorhabditis elegans paramyosin: thus, λcDi2 encodes D. immitis paramyosin.

AB - The cDNA synthesized from mRNA of Dirofilaria immitis female adult worms was cloned into the expression vector λgt11. Screening the library with a hyperimmune rabbit antiserum raised against adult worm homogenates yielded several antigen positive clones. One of these clones, λcDi2, was recognized by rabbit antisera raised against either D. immitis L-3, adult, Brugia malayi L-3 or Onchocerca volvulus adult worm antigen, as well as by antisera from humans naturally infected with O. volvulus or Wuchereria bancrofti. Affinity-purified anti-λcDi2 antibodies reacted with a 97-kDa protein on Western transfers of adult D. immitis antigen extracts that were reduced with β-mercaptoethanol. The whole rabbit anti-D. immitis adult antiserum depleted of anti-λcDi2 antibodies exhibited decreased reactivity to this 97-kDa band. A monoclonal antibody (IA6) that specifically binds Schistosoma mansoni paramyosin also recognised a 97-kDa protein in D. immitis extracts upon Western transfer. The deduced amino acid sequence of partial DNA sequence from λcDi2 showed some similarity to nematode myosin, and gave a stretch of 82 amino acids that is 91.5% identical to Caenorhabditis elegans paramyosin: thus, λcDi2 encodes D. immitis paramyosin.

KW - Affinity-purified antibody

KW - Dirofilaria immitis

KW - Myosin

KW - Paramyosin

KW - λgt11 cDNA library

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DO - 10.1016/0166-6851(89)90139-4

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C2 - 2527335

AN - SCOPUS:0024318833

SN - 0166-6851

VL - 35

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EP - 41

JO - Molecular and Biochemical Parasitology

JF - Molecular and Biochemical Parasitology

IS - 1

ER -

A λgt11 cDNA recombinant that encodes Dirofilaria immitis paramyosin

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Keywords

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