Co-expression Of PCa05 And EGFP In LNCaP Cells And Its Subcellular Localization

Prostatic cancer is one of the most common cancer and death cause among men in the world especially in Euramerican countries. With the aging of population and development of examination,its incidence is increased year by year.In terms of the statistics of prostatic cancer incidence,it is ranked one of the fastest increased cancer in the ,21^st century in our country.In the past experiment,we found that gene PCaO5 was higher expressed in the expression database of the prostatic cancer tissue,its expression was 10 times higher than that in the common prostatic tissue.On the other hand, the expression of gene PCaO5 in the prostatic cancer tissue was higher than that in the other cancerous and common tissue.The result of Northern Blot originally indicated the upregulation of gene PCaO5 expression in the prostatic cancer. On the other hand, we did not find any report of this gene.According to PCaO5 sequence, a pair of primers containing the sites for given restrictive endonuclease at both ends were designed and synthesized. RT-PCR of the total RNA extracted from LNCaP cell was performed, then abtained the objective PCaO5 gene segment. By digesting the eukaryotic expression vector pEGFP-C1 and objective PCaO5 gene segment with two types of restrictively nucleic acid inscribed enzyme Hind III and EcoR I, we obtained linearization pEGFP-C1 vector and objective PCaO5 gene segment, and joined them together, consequently gained recombined plasmid pEGFP-C1-PCaO5. The recombinants were sequenced and identified by restrictive endonuclease digestion and then transfected into the LNCaP cell by Lipofectamine 2000 and we observed their subcellular localization of pEGFP-C1-PCaO5 by the Laser scannning confocal microscope after 24 hour cell creep parcel.Some useful predicted information was obtained about gene function of PCa05, and PCaO5 is possibly a soluble protein regulated by oct-1 and revolved in regulating the gene expressing in celluar mitochondrion. The fusion protein was expressed in both nuclei and cytoplasm of LNCaP cell. However, compared with pure EGFP, the expression of...