title: Construction of cytoplasmic molecular markers distinguishing Danio rerio from Gobiocypris rarus at high identity domains based on MP-PCR strategy and Sybr Green I detection.
abstract: To distinguish the cytoplasm of Danio rerio from that of Gobiocypris rarus, we cloned G. rarus COXI and constructed cytoplasmic molecular markers at the high identity domains of COXI by mutated primer PCR (MP-PCR for short). Then Sybr Green I was used to detect the single amplicon. As a result, we succeeded in getting the cytoplasmic molecular markers, G.M COXI and Z.M COXI, by MP-PCR strategy. They were used to detect the sperm-derived mtDNA in the sexual hybrid embryos (D. rerio female symbolxG. rarus male symbol) before the sphere stage. In the present study, all results demonstrate that MP-PCR approach and Sybr Green I detection are feasible to construct the molecular markers to identify genes that shared high identity.
authors.affiliation: State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China.
authors.forename: De-Sheng
authors.initials: DS
authors.lastname: Pei
authors.forename: Yong-Hua
authors.initials: YH
authors.lastname: Sun
authors.forename: Zuo-Yan
authors.initials: ZY
authors.lastname: Zhu
mesh_terms: D000818:Animals; D001483:Base Sequence; D052160:Benzothiazoles; D015415:Biomarkers; D003001:Cloning, Molecular; D051545:Cyclooxygenase 1 / Q000378:metabolism*; D003530:Cyprinidae / Q000378:metabolism*; D003593:Cytoplasm / Q000201:enzymology*; D017931:DNA Primers / Q000378:metabolism; D004272:DNA, Mitochondrial / Q000378:metabolism; D003959:Diamines; D004587:Electrophoresis, Agar Gel; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D006824:Hybridization, Genetic; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009930:Organic Chemicals / Q000378:metabolism*; D010063:Ovum / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011804:Quinolines; D016415:Sequence Alignment; D013094:Spermatozoa / Q000378:metabolism; D015027:Zebrafish / Q000378:metabolism*
chemical_list: D052160:Benzothiazoles; D015415:Biomarkers; D017931:DNA Primers; D004272:DNA, Mitochondrial; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D051545:Cyclooxygenase 1