[Preparation of RNA probe for cd99l2 gene of zebrafish labeled with digoxingenin-UTP].
OBJECTIVE
To study the expression pattern of cd99l2 gene during zebrafish development, the RNA probes for whole-mount in situ hybridization were prepared in this study.


METHODS
The 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.


RESULTS
The 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.


CONCLUSIONS
The antisense probe can be used for study of the spatial and temporal distribution of cd99l2 during zebrafish development using the sense probe as control.
Wen
Zong-hua
ZH
Department of Pathology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. wenzonghua@126.com
Zhang
Yan
Y
Wu
Zi-qin
ZQ
Zhou
Xin-hua
XH
Han
Xi-qun
XQ
Zhang
Wen-qing
WQ
Zhao
Tong
T
D000818:Animals; D002490:Central Nervous System / Q000196:embryology; D003001:Cloning, Molecular; D004076:Digoxigenin / Q000737:chemistry; D018507:Gene Expression Regulation, Developmental; D015345:Oligonucleotide Probes; D015347:RNA Probes; D014544:Uridine Triphosphate / Q000737:chemistry; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics*
D015345:Oligonucleotide Probes; D015347:RNA Probes; D029961:Zebrafish Proteins; D004076:Digoxigenin; D014544:Uridine Triphosphate