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.
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.
Pei
De-Sheng
DS
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China.
Sun
Yong-Hua
YH
Zhu
Zuo-Yan
ZY
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*
D052160:Benzothiazoles; D015415:Biomarkers; D017931:DNA Primers; D004272:DNA, Mitochondrial; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I; D051545:Cyclooxygenase 1