[Cloning and subcellular localization of apr-1--a new gene of tumor specific antigen family].
OBJECTIVE
apr-1 was cloned by improved polymerase chain reaction (PCR)-based subtractive hybridization from all-trans retinoic acid (ATRA)-induced apoptotic leukemia HL-60 cells in 1999. Preliminary results showed that apr-1 might be an apoptosis-related gene (GenBank ID: NM_014061). This study was to explore the background of apr-1 through gene cloning, bioinformatic analysis, and subcellular locating.


METHODS
The cDNA encoding Apr-1 was amplified by reverse transcription-PCR (RT-PCR), and sequenced. Open reading frame (ORF) of apr-1 was analyzed with ORF finder software. Chromosome locus was defined by genome blast software. Conserved domains of amino acids were analyzed by protein blast software. Align (Cluster W) software in Vector NTI software package was used to analyze homogeneous genes (or proteins), and to draw the Phylogenetic Tree. Subcellular localization of apr-1 was performed.


RESULTS
apr-1 was mapped to chromosome Xp11.22 with the ORF locating in 1 exon. Two MAGE conserved domains were found in Apr-1. Apr-1 shared homology with MAGE-A1, MAGE-B1, MAGE-C1, MAGE-D1, and Necdin. Phylogenetic analysis showed that Apr-1 was more closely related to MAGE-D1 and Necdin. Gene products of apr-1 were located in the nuclei of eukaryocytes.


CONCLUSIONS
apr-1 is a member of MAGE family, and might belong to type II MAGE genes.
Yan
Wei
W
Department of Pathology, Xijing Hospital, The Fourth Military Medical University, Xi'an, Shaanxi, 710032, P. R. China. yan070@sohu.com
Wang
Wen-Liang
WL
Zhu
Feng
F
Cheng
Sheng-Quan
SQ
Li
Qin-Long
QL
Wang
Li
L
Wang
Cheng-Ji
CJ
D000595:Amino Acid Sequence; D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics*; D001483:Base Sequence; D019556:COS Cells; D002467:Cell Nucleus / Q000737:chemistry; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics*; D018922:HL-60 Cells; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000032:analysis / Q000235:genetics*; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000032:analysis / Q000235:genetics*; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics*; D016366:Open Reading Frames; D010802:Phylogeny; D016415:Sequence Alignment; D014960:X Chromosome
D000951:Antigens, Neoplasm; D018076:DNA, Complementary; C120897:MAGED1 protein, human; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; C069766:necdin; C076744:cytoplasmic linker protein 170