Isolation of cDNA encoding immunoglobulin light chain from common carp (Cyprinus carpio L.)

Mitsuru Tomana, Miki Nakao, Tadaaki Moritomo, Kazuhiro Fujiki, Tomoki Yano

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

cDNA clones partially encoding immunoglobulin (lg) light (L) chain variable (V) region associated with constant (C) region were isolated from carp (Cyprinus carpio L.) spleen by reverse transcription-polymerase chain reaction (RT-PCR) using degenerated primers corresponding to the conserved amino acid sequences in N-terminus of VL domain and in middle of CL domain. The VL region is readily delineated into framework regions (FR) and complementarity-determining regions (CDR). cDNA clones encoding complete CL domain were then obtained by a rapid amplification of cDNA 3′-end method (3′-RACE). The sequences of both VL and CL region contain well conserved cysteine residues important for intra- or interdomain interaction in mammals. Comparisons of the amino acid sequence of the CL domain with those of other species showed the highest degree of similarity of 59%, 55%, and 54% to those of rainbow trout IgL 1 isotype (Oncorhynchus mykiss), channel catfish G isotype (Ictalurus punctatus), and Atlantic cod (Gadus morhua L.), respectively. Multiple sequence alignments of the CL domain with those of higher vertebrate, however, did not readily allow it to be classified as κ or λ isotypes. Genomic Southern blot analyses using VL-and CL-specific probes showed that there are at least 13 VL segments and 14 CL segments in the genome of the carp, indicating that IgL locus of carp has a cluster-like organisation similar to those of several teleost species.

Original languageEnglish
Pages (from-to)71-80
Number of pages10
JournalFish and Shellfish Immunology
Volume9
Issue number2
DOIs
Publication statusPublished - Jan 1 1999

Fingerprint

immunoglobulin light chains
Immunoglobulin Light Chains
Cyprinus carpio
carp
Complementary DNA
Ictalurus punctatus
Gadus morhua
Oncorhynchus mykiss
amino acid sequences
clones
Complementarity Determining Regions
Amino Acids
clone
Mammals
amino acid
Polymerase chain reaction
sequence alignment
Transcription
Southern blotting
immunoglobulins

All Science Journal Classification (ASJC) codes

  • Environmental Chemistry
  • Aquatic Science

Cite this

Isolation of cDNA encoding immunoglobulin light chain from common carp (Cyprinus carpio L.). / Tomana, Mitsuru; Nakao, Miki; Moritomo, Tadaaki; Fujiki, Kazuhiro; Yano, Tomoki.

In: Fish and Shellfish Immunology, Vol. 9, No. 2, 01.01.1999, p. 71-80.

Research output: Contribution to journalArticle

Tomana, Mitsuru ; Nakao, Miki ; Moritomo, Tadaaki ; Fujiki, Kazuhiro ; Yano, Tomoki. / Isolation of cDNA encoding immunoglobulin light chain from common carp (Cyprinus carpio L.). In: Fish and Shellfish Immunology. 1999 ; Vol. 9, No. 2. pp. 71-80.
@article{28ac9eef0f7a44159eacccc8bc039a9e,
title = "Isolation of cDNA encoding immunoglobulin light chain from common carp (Cyprinus carpio L.)",
abstract = "cDNA clones partially encoding immunoglobulin (lg) light (L) chain variable (V) region associated with constant (C) region were isolated from carp (Cyprinus carpio L.) spleen by reverse transcription-polymerase chain reaction (RT-PCR) using degenerated primers corresponding to the conserved amino acid sequences in N-terminus of VL domain and in middle of CL domain. The VL region is readily delineated into framework regions (FR) and complementarity-determining regions (CDR). cDNA clones encoding complete CL domain were then obtained by a rapid amplification of cDNA 3′-end method (3′-RACE). The sequences of both VL and CL region contain well conserved cysteine residues important for intra- or interdomain interaction in mammals. Comparisons of the amino acid sequence of the CL domain with those of other species showed the highest degree of similarity of 59{\%}, 55{\%}, and 54{\%} to those of rainbow trout IgL 1 isotype (Oncorhynchus mykiss), channel catfish G isotype (Ictalurus punctatus), and Atlantic cod (Gadus morhua L.), respectively. Multiple sequence alignments of the CL domain with those of higher vertebrate, however, did not readily allow it to be classified as κ or λ isotypes. Genomic Southern blot analyses using VL-and CL-specific probes showed that there are at least 13 VL segments and 14 CL segments in the genome of the carp, indicating that IgL locus of carp has a cluster-like organisation similar to those of several teleost species.",
author = "Mitsuru Tomana and Miki Nakao and Tadaaki Moritomo and Kazuhiro Fujiki and Tomoki Yano",
year = "1999",
month = "1",
day = "1",
doi = "10.1006/fsim.1998.0175",
language = "English",
volume = "9",
pages = "71--80",
journal = "Fish and Shellfish Immunology",
issn = "1050-4648",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Isolation of cDNA encoding immunoglobulin light chain from common carp (Cyprinus carpio L.)

AU - Tomana, Mitsuru

AU - Nakao, Miki

AU - Moritomo, Tadaaki

AU - Fujiki, Kazuhiro

AU - Yano, Tomoki

PY - 1999/1/1

Y1 - 1999/1/1

N2 - cDNA clones partially encoding immunoglobulin (lg) light (L) chain variable (V) region associated with constant (C) region were isolated from carp (Cyprinus carpio L.) spleen by reverse transcription-polymerase chain reaction (RT-PCR) using degenerated primers corresponding to the conserved amino acid sequences in N-terminus of VL domain and in middle of CL domain. The VL region is readily delineated into framework regions (FR) and complementarity-determining regions (CDR). cDNA clones encoding complete CL domain were then obtained by a rapid amplification of cDNA 3′-end method (3′-RACE). The sequences of both VL and CL region contain well conserved cysteine residues important for intra- or interdomain interaction in mammals. Comparisons of the amino acid sequence of the CL domain with those of other species showed the highest degree of similarity of 59%, 55%, and 54% to those of rainbow trout IgL 1 isotype (Oncorhynchus mykiss), channel catfish G isotype (Ictalurus punctatus), and Atlantic cod (Gadus morhua L.), respectively. Multiple sequence alignments of the CL domain with those of higher vertebrate, however, did not readily allow it to be classified as κ or λ isotypes. Genomic Southern blot analyses using VL-and CL-specific probes showed that there are at least 13 VL segments and 14 CL segments in the genome of the carp, indicating that IgL locus of carp has a cluster-like organisation similar to those of several teleost species.

AB - cDNA clones partially encoding immunoglobulin (lg) light (L) chain variable (V) region associated with constant (C) region were isolated from carp (Cyprinus carpio L.) spleen by reverse transcription-polymerase chain reaction (RT-PCR) using degenerated primers corresponding to the conserved amino acid sequences in N-terminus of VL domain and in middle of CL domain. The VL region is readily delineated into framework regions (FR) and complementarity-determining regions (CDR). cDNA clones encoding complete CL domain were then obtained by a rapid amplification of cDNA 3′-end method (3′-RACE). The sequences of both VL and CL region contain well conserved cysteine residues important for intra- or interdomain interaction in mammals. Comparisons of the amino acid sequence of the CL domain with those of other species showed the highest degree of similarity of 59%, 55%, and 54% to those of rainbow trout IgL 1 isotype (Oncorhynchus mykiss), channel catfish G isotype (Ictalurus punctatus), and Atlantic cod (Gadus morhua L.), respectively. Multiple sequence alignments of the CL domain with those of higher vertebrate, however, did not readily allow it to be classified as κ or λ isotypes. Genomic Southern blot analyses using VL-and CL-specific probes showed that there are at least 13 VL segments and 14 CL segments in the genome of the carp, indicating that IgL locus of carp has a cluster-like organisation similar to those of several teleost species.

UR - http://www.scopus.com/inward/record.url?scp=0033074577&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033074577&partnerID=8YFLogxK

U2 - 10.1006/fsim.1998.0175

DO - 10.1006/fsim.1998.0175

M3 - Article

AN - SCOPUS:0033074577

VL - 9

SP - 71

EP - 80

JO - Fish and Shellfish Immunology

JF - Fish and Shellfish Immunology

SN - 1050-4648

IS - 2

ER -