[Cloning, sequencing and bioinformatics analysis of a new tumor suppressor gene ndr2 from mouse].
OBJECTIVE
Ndr2 (N-myc down stream regulator) gene in human is a new gene cloned with the human adult whole brain cDNA as template in 1999, which accession number is AF159092 in GenBank. Locating backward position of the N-myc gene in human chromosome, this gene was named Ndr2 gene. The previous experimental results showed Ndr2 gene probably is a tumor suppressor gene. To research the function of Ndr2 gene, the authors cloned the genomic sequence of ndr2 from mouse.


METHODS
To clone Ndr2 genomic sequence by reverse transcription-polymerase chain reaction(RT-PCR) with the mouse genome library as template; automatic sequencing was performed using 310 Genetic Analyzer; homogeneous analysis was made using GenBank BLAST; open reading fragment(ORF) analysis was made using PC Gene and ORF Finder; domain analysis was made using ProDom system.


RESULTS
A fragment (about 3310bp,identified by agarose gel electrophoresis) was obtained using RT-PCR with the mouse genome library as template. The fragment was cloned in pMD18-T vector. BLAST analysis showed that the sequence was highly homogeneous (with the homogeneity rate of 91.4%) with Ndr2 gene in human and non-homogeneous with genomic sequence database in mouse. ORF analysis showed that there was a complete coding region in it, which including 8 extrons and 7 introns; it can interpret a protein containing about 200 amino acid residuals. ProDom analysis showed there was a domain like acyl carrier protein(ACP) in it.


CONCLUSIONS
The authors cloned Ndr2 gene in mouse and proved that the sequence is a new genome sequence in mouse genomic sequence database. At present, the genome sequence has been submitted to GenBank(the accession number: AY151387).
Yang
Xing-Long
XL
Department of Biochemistry and Molecular Biology, The Fourth Military Medical University, Xi'an, Shanxi, 710032, P.R.China.
Zhang
Ya-Li
YL
Yao
Li-Bo
LB
Liu
Xin-Ping
XP
Ji
Shao-Ping
SP
Xing
Fei-Yue
FY
D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D019295:Computational Biology; D004247:DNA / Q000032:analysis; D016147:Genes, Tumor Suppressor; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D055456:Nodal Signaling Ligands; D011506:Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000235:genetics*
D048868:Adaptor Proteins, Signal Transducing; C485172:Ndr2 protein, mouse; C113775:Ndr2 protein, vertebrate; D055456:Nodal Signaling Ligands; D011506:Proteins; D016212:Transforming Growth Factor beta; D004247:DNA