Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevis

Two female-specific AFLP (amplified fragment length polymorphism) markers (named CseF464 and CseF136) were isolated by using one selective primer combination (E-AGC/M-CTG) from the genomic DNA of 20 females and 20 males of the half-smooth tongue sole Cynoglossus semilaevis. Both the markers were re-...

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Main Author: Hongyu MA, Songlin CHEN, Jing LI, Jin-Zhen BI, Tianjun XU
Format: Article
Language:English
Published: Oxford University Press 2009-08-01
Series:Current Zoology
Subjects:
Online Access:http://www.currentzoology.org/paperdetail.asp?id=11230
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spelling doaj-9a5d01efaf0e44bc9cd1aab32cc2a1a12020-11-24T23:22:31ZengOxford University PressCurrent Zoology1674-55072009-08-01554309314Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevisHongyu MA, Songlin CHEN, Jing LI, Jin-Zhen BI, Tianjun XUTwo female-specific AFLP (amplified fragment length polymorphism) markers (named CseF464 and CseF136) were isolated by using one selective primer combination (E-AGC/M-CTG) from the genomic DNA of 20 females and 20 males of the half-smooth tongue sole Cynoglossus semilaevis. Both the markers were re-amplified, recovered from the agarose gels, cloned and sequenced. Bioinformatics analysis indicated that the length of the two markers were 468bp and 134bp, respectively, and the sequences showed no similarity to each other, as well as to the known sequences deposited in the GenBank database using BLASTn. Two pairs of SCAR (sequence characterized amplified regions) primers were designed based on the sequences of the two female-specific markers. Furthermore, PCR-based genetic sex identification method was developed in Cynoglossus semilaevis. A specific fragment was amplified in all females but not in any males by using these SCAR primers on the initial 20 female and 20 male individuals of Cynoglossus semilaevis. The feasibility of the two SCAR primer pairs was confirmed in additional 100 individuals (50 females and 50 males). This allowed for reliable, rapid molecular identification of genetic sex of the species, genetic mapping on the sex chromosomes and better understanding of the sex determination and sex differentiation in the half-smooth tongue sole [Current Zoology 55 (4): 309–314, 2009]. http://www.currentzoology.org/paperdetail.asp?id=11230AFLPCynoglossus semilaevisFemale-specific markerSCARMolecular sexing
collection DOAJ
language English
format Article
sources DOAJ
author Hongyu MA, Songlin CHEN, Jing LI, Jin-Zhen BI, Tianjun XU
spellingShingle Hongyu MA, Songlin CHEN, Jing LI, Jin-Zhen BI, Tianjun XU
Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevis
Current Zoology
AFLP
Cynoglossus semilaevis
Female-specific marker
SCAR
Molecular sexing
author_facet Hongyu MA, Songlin CHEN, Jing LI, Jin-Zhen BI, Tianjun XU
author_sort Hongyu MA, Songlin CHEN, Jing LI, Jin-Zhen BI, Tianjun XU
title Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevis
title_short Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevis
title_full Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevis
title_fullStr Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevis
title_full_unstemmed Cloning, characterization of two female-specific AFLP markers and development of PCR-based sex identification method for the half-smooth tongue sole Cynoglossus semilaevis
title_sort cloning, characterization of two female-specific aflp markers and development of pcr-based sex identification method for the half-smooth tongue sole cynoglossus semilaevis
publisher Oxford University Press
series Current Zoology
issn 1674-5507
publishDate 2009-08-01
description Two female-specific AFLP (amplified fragment length polymorphism) markers (named CseF464 and CseF136) were isolated by using one selective primer combination (E-AGC/M-CTG) from the genomic DNA of 20 females and 20 males of the half-smooth tongue sole Cynoglossus semilaevis. Both the markers were re-amplified, recovered from the agarose gels, cloned and sequenced. Bioinformatics analysis indicated that the length of the two markers were 468bp and 134bp, respectively, and the sequences showed no similarity to each other, as well as to the known sequences deposited in the GenBank database using BLASTn. Two pairs of SCAR (sequence characterized amplified regions) primers were designed based on the sequences of the two female-specific markers. Furthermore, PCR-based genetic sex identification method was developed in Cynoglossus semilaevis. A specific fragment was amplified in all females but not in any males by using these SCAR primers on the initial 20 female and 20 male individuals of Cynoglossus semilaevis. The feasibility of the two SCAR primer pairs was confirmed in additional 100 individuals (50 females and 50 males). This allowed for reliable, rapid molecular identification of genetic sex of the species, genetic mapping on the sex chromosomes and better understanding of the sex determination and sex differentiation in the half-smooth tongue sole [Current Zoology 55 (4): 309–314, 2009].
topic AFLP
Cynoglossus semilaevis
Female-specific marker
SCAR
Molecular sexing
url http://www.currentzoology.org/paperdetail.asp?id=11230
work_keys_str_mv AT hongyumasonglinchenjinglijinzhenbitianjunxu cloningcharacterizationoftwofemalespecificaflpmarkersanddevelopmentofpcrbasedsexidentificationmethodforthehalfsmoothtonguesolecynoglossussemilaevis
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