Objective: To investigate the molecular cloning, tissue distribution, tumor distribution and the subcellular localization of AK078438 gene.
Methods: Reverse transcriptase-polymerase chain reaction (RT-PCR) technique was applied to amplify the open reading frame of AK078438 gene. Prokaryotic expressing vector and affinity purification were used to get the fusion protein. At mRNA expressing levels semi-quantitative RT-PCR was employed for the investigation of its distribution in normal rat tissues, tumor cell and mesenchymal stem cell (MSC) lines. The eukaryotic expression green fluorescence protein (GFP) fusion protein vector was transfected into cells and identified subcellular localization.
Results: The full open reading frame of AK078438 gene with 1065 bp, 355aa were identified. mRNA of the gene distributed in brain, uterus, colon, bone marrow, testis and kidney, but not in stomach, liver, the lung, spleen and heart. Murine colon adenocarcinoma C26, melanoma B16, Lewis lung carcinoma LL/2 and MSC had the gene and mouse myeloma cell line NS-1 and Hepa mouse hepatoma cell line had no the gene. The protein was localized in cytoplasm.
Conclusion: Protein expression, expressing profile and subcellular localization of AK078438 gene set the basis for further exploration of its functions.