Preparation of RNA probe for cd99l2 gene of zebrafish labeled with digoxingenin-UTP.
- Author:
Zong-hua WEN
1
;
Yan ZHANG
;
Zi-qin WU
;
Xin-hua ZHOU
;
Xi-qun HAN
;
Wen-qing ZHANG
;
Tong ZHAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Central Nervous System; embryology; Cloning, Molecular; Digoxigenin; chemistry; Gene Expression Regulation, Developmental; Oligonucleotide Probes; RNA Probes; Uridine Triphosphate; chemistry; Zebrafish; embryology; genetics; Zebrafish Proteins; genetics
- From: Journal of Southern Medical University 2010;30(5):969-972
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the expression pattern of cd99l2 gene during zebrafish development, the RNA probes for whole-mount in situ hybridization were prepared in this study.
METHODSThe cd99l2 fragment obtained by RT-PCR was cloned into pGM-T Easy, then the plasmids were linearized with the restriction enzymes SacII or SalI. Using Sp6 or T(7) RNA polymerase, the digoxingenin-labeled antisense and sense probes were synthesized and confirmed by whole-mount in situ hybridization.
RESULTSThe plasmid cd99l2/pGM-T was constructed. cd99l2 gene expression pattern during embryogenesis of zebrafish was examined using the antisense probe, and intense expression was detected in the central nervous system during zebrafish development.
CONCLUSIONThe antisense probe can be used for study of the spatial and temporal distribution of cd99l2 during zebrafish development using the sense probe as control.
