Content uploaded by Michel Leclerc
Author content
All content in this area was uploaded by Michel Leclerc on Feb 14, 2018
Content may be subject to copyright.
International Journal of Biotechnology and Bioengineering Volume 1 Issue 2, September 2016
Michel Leclerc et al, IJBB 2016, 1:2
International Journal of Biotechnology and
Bioengineering
Evidence of Fab Fragment gene in an Invertebrate : the sea star Asterias rubens
Research Article Open Access
Michel Leclerc1*, Nicolas Kresdorn2
1556 rue Isabelle Romée Sandillon, France
2 GenXpro de , Frankfurt, Germany
*Corresponding Author: Michel Leclerc, 556 rue Isabelle Romée Sandillon,France. Email : mleclerc45@gmail.com
Citation: Michel Leclerc, Nicolas Kresdorn (2016) Evidence of Fab Fragment gene in an Invertebrate : the sea star Asterias rubens.
Int J Biotech & Bioeng. 1:2, 52-53
Copyright: © 2016 Michel Leclerc et al. This is an open-access article distributed under the terms of the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are
credited..
Abstract
Fc receptor gene,, was just discovered with the emergence of Fab fragments gene, in Asterias rubens These last ones were
described in Ig light chain and Ig heavy chain , when compared to mouse genome.
52
Introduction
The purpose of this article is to draw attention to the mass of
information that has accumulated on the primitive sea star
antibody since about 1983. This work has emanated from our
laboratory, those of Gothenburg (Dr Sam Dupont), those of GenX
pro (Frankfurt),those of Fasteris (Switzerland).
Our purpose will be to recall, the homologies between the
primitive antibody and the mouse Igkappa chain [1],the presence
of two Immunoglobulin sites in this last one, the specicity
of this « antibody » to the the antigen HRP which induces the
reaction : »antigen-antibody », at last the existence in the sea star
immune system of the Fc receptor gene [2] We present now, the
last results, concerning this primitive antibody : the emergence
of Fab fragments in the immunized and non-immunized sea star
transcriptomes, when compared to mouse genome.
Materials and Methods
Sea stars were obtained from the Biology Institute (Gothenburg
University) Immunizations to HRP (Horse-radish peroxydase), ge-
nomic studies were already described [1]. Briey, after ligation of
adapters for Illumina’s GSII sequencing system, the cDNA was se-
quenced on the Illumina GSII platform sequencing. 1.100 bp from
one side of the approximately 200 bp fragments sequences were
assembled using Velvet (3)
Results
First a comparison between non-immunized sea star genome and
mouse genome was performed.Studies were compared to mouse
Immunoglobulin Light chain, then to heavy chain :
One contig (Contig9579) could be annotated via TBLASTX
to Mus musculus “Mus musculus immunoglobulin IgG2a light
chain Fab fragment (Mab10CK) mRNA, partial cds” from the
NCBI database (gi|2852986|gb|AF042656.1|), with an e-value of
4.15e-05. On an aligned region of 21 amino acids, 14 positive and
9 identical amino acids were found.
5’CAGAGTCACCAGGTGCTGTGTTGACATCAG-
CACTTTCTGAATCCACCTCCATTGGTTGAT
CCACTTCCTCTTTCACTATAGTTGACGATTT-
GATATCAACACTGTCACCTGTAAGCGCAT
CAACACCGCTTGTTATATCAACAGAC-
GATCTCCTTTGTGGATCCACTTTTTGATCGGTGG
ATAGAGGCGGAGTGGATGACTCCATCTTTGCTA-
ATGTTTTGTCATCGGCTAATGTTGATG
AACTTTCTTCAGCTTTTACCTCGGTCTTCA-
CATCCACTGTTTGGTTTTCTGATTTCACGG
AAACAGTTGATGTCGTTTTGACTGGATC-
CACTGACTTATCAGCCAATGTTCCCTGCGGAG
TTGTTGATGTACTTTGTTGAGAGATGGCTGT-
GAAGGGATTTGTGGCGGGATCGAAGGAAG
Received August 23, 2016; Accepted September 19, 2016; Published September 29, 2016.
International Journal of Biotechnology and Bioengineering Volume 1 Issue 2, September 2016
Michel Leclerc et al, IJBB 2016, 1:2
53
CTCCTAAGCCTTCCTCCCCTGGGGTTGGC-
GTTTCTAGAAGCGAGGAATCTTTGGCATCTT
CGAAACACAAGGGATGGTATGTCCTGGAGTT-
GAACCTGACAGCATCTCTCAGTTTCCAAA
CCTCTTCATCTTCATCAAAGAATTGCTCAAAC-
GGCTCATGACACAGAGAACACATCTCAT
CGCCAGTGACGTTTCCATCAATAGCCACA-
CAGAGGATGCCATCTTTATCAGGTTTAGACT
CTGCAGTCATCTCCTCAAAGAAACT-
GCTCCTCTCTTCATCCACTTCGTTGATTTCTTCAA
AGTCCAACCACTGATCGACTTCATAGAAC-
CACTTCCTGGACTGAACAGCCTTCACTCCAT
CCTTCTCACGACGATTCTGACGGAAGTGC-
CAGTCCAGATGTTTCCGATATCTGTCCATTT
CATCAATGACAAAGCGCATACCA3’
Second, results with mouse Ig heavy chain were obtained, when
compared to mouse genome : One contig (c37102_g1_i1) could be
annotated via TBLASTX to Mus musculus “Mus musculus immu-
noglobulin IgG2a heavy chain Fab fragment (Mab10CH) mRNA,
partial cds” from the NCBI database (gi|2852988|gb|AF042657.1|),
with an e-value of 6.1e-06. On an aligned region of 20 amino ac-
ids, 14 positive and 10 identical amino acids were found.
5’AACTGCGCCACCAGCAAGTATGCCGTAC-
CCTTCGGCTATTGCGACGGCACCAGACTACCC
CAAAGCAATGGCAGCATCGGCCCCTCCATTTG-
GTGGTGAAGCTCCGCCCCCAAGTTACGA
GGAGGCTGTTGGCAAAACAGACAGACTGG-
CCAACGCAAGGAACGATGAGGATTACGTCAT
TGACGATCAGTTCGTGCCTAGTTATCCG-
TACTTCAACCTTTCGGTTGATGTACCACCGGG
TTCCCGCCATTAACCCCATGGAGTTTAAAGAGA3’
At last, results with immunized sea star genome were given
when compared to mouse IgG2a light chain : One contig (Con-
tig5278|m.6828) could be annotated via TBLASTX to Mus mus-
culus “Mus musculus immunoglobulin IgG2a light chain Fab frag-
ment (Mab10CK) mRNA, partial cds” from the NCBI database
(gi|2852986|gb|AF042656.1|), with an e-value of 1.62e-05. On an
aligned region of 38 amino acids, 23 positive and 14 identical ami-
no acids were found.
5’TCTTTGGAAACCCCGAACCGGAACTGACCT-
GGCAGAGGAAGAGCCCAGCAGGTGCAGACC
TACCACTCGGTGCAGAGCTCCGATTCAA-
CAACCAGCATTTGATCCTCAGGAATATACAGA
TTGAAGATGCTGGGGAGTATGAATGTATGAC-
CACTAATGGTGGAGGAAGAACATCTGGAA
CCATTACTGTCCGAGTCCCACCCACCTGGCT-
GCAAGAAATCTCTGACATCGAGGGGAACA
TCTACGGAGACTTATCCATGAAGTGTAAAG-
CAGAGAGTCCCGATGGCGTAACCTACACAT
GGTACCGCAATGCTGAGGTGATCCAAGACCTTC-
CCAGGCATCAGTTCAGCAACGGTTTGA
AGACACTCACCATCCAGAATCTGACTGTG-
GAAGACACCGGGATGTACCAGTGTATGGCTA
GTAATGAATACGGCCAGATATACACCACAG-
CACAGGTCACTGTACAAGCCATTGCACCAT
GGTTTGAGACGCCCCTTGAGGTGGACCAGC-
CAGCACCCCGTGGTGGCTCAGTGACCATCC
TCTGCCAACCTAAAGCTGCACCCACACCCGT-
CATCGTCTGGACCAAAGGAAGCGAGACCA
TTGAGAACGGTGGCCGCTACACCGTCATG-
GACAACGGTGATTTGATGATCAGTGGTGTTA
GTGATGCAGATGGAGGAGACTACACCTG-
TACGGCTACCAATAGCATGGGAACTGATTCAA
GCACTGGTAGCTTGATCATCAAGGATGGTA-
CAGCCATGGTGGAGCCACCCATGGCATTGA
CTGTAAACGAAGGGGAGATGGGCACATT-
GACCTGCCGTGCATCACACGATCCGTTCTTTG
AGCTGGAGTATGTGTGGCTGAAAGAAG-
GAATCCAAGTTGATCCTCGTCTCCCTAACTATG
AGATACCTCAGATCCAAATGGGCGAGCCAAGT-
CAGCTGCATATCAAAGACGCCACTATGG
ATCTAGGAGGGGAGTACATGTGC3’
Discussion and Conclusion
It is clear that mouse Fab IgG2a light chain gene, mouse Fab Ig-
G2a heavy chain gene, are present in the sea star genome. It is
also noticeable that these genes ,show a « symmetry », or at least
a similarity in their congurations : we count on an aligned region
of 21 amino-acids , 14 positive and 9 identical in the case of IgG2a
light chain, 14 positive and 10 identical in the case of IgG2a heavy
chain out of 20 amino-acids.
As for immunized sea star IgG2a light chain which is only studied,
in the present work : the number of positive amino-acids increases,
in a signicant manner (on an aligned region of 38 amino-acids, 23
positive instead 9 in non-immunized sea stars to HRP : undoubtly
it is due to immunization( further studies are bound to unravel the
phenomenon). In addition to the work on Fc receptor gene [2],
it appears that Fab fragments gene, corroborate the existence of
the Invertebrate Primitive Antibody through the Asterias rubens
history [4].
References
1. Vincent N, et al, Metagene 2(2014) 320-322
2. Leclerc M., et al Immunol. Lett (2016) submitted
3. Zerbino DR., et al, Gen Res (2007) 18:821-9
4. Leclerc M, « Sea star Genomic studies Following 2 » (2016)
Lambert Ed. (Germany