ZFIN ID: ZDB-PUB-131031-1
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
Pei, D.S., Sun, Y.H., and Zhu, Z.Y.
Date: 2008
Source: Molecular biology reports   35(1): 45-50 (Journal)
Registered Authors: Pei, Desheng, Sun, Yonghua, Zhu, Zuoyan
Keywords: COXI, Danio rerio, Gobiocypris rarus, MP-PCR, Sybr Green | detection
MeSH Terms:
  • Animals
  • Base Sequence
  • Biomarkers
  • Cloning, Molecular
  • Cyclooxygenase 1/metabolism*
  • Cyprinidae/metabolism*
  • Cytoplasm/enzymology*
  • DNA Primers/metabolism
  • DNA, Mitochondrial/metabolism
  • Electrophoresis, Agar Gel
  • Embryo, Nonmammalian/metabolism
  • Female
  • Hybridization, Genetic
  • Male
  • Molecular Sequence Data
  • Mutation/genetics
  • Organic Chemicals/metabolism*
  • Ovum/metabolism
  • Polymerase Chain Reaction/methods*
  • Sequence Alignment
  • Spermatozoa/metabolism
  • Zebrafish/metabolism*
PubMed: 17211516 Full text @ Mol. Biol. Rep.

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 × G. rarus male) 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.