ABSTRACT
This work is aimed at the investigation of the senescence associated genes differentially expressed in rat muscle by cDNA microarray. After total RNA isolation, mRNA purification and probe preparation, the gene expression patterns of rat muscle from the old group (30 months old) and the young group (3 months old) were compared by cDNA gene chips. In the muscle of old rats there were reproducibly 127 differentially expressed genes, among which some down-regulated genes were involved in energy metabolism and signal transduction, while some up-regulated genes were related to protein degradation and cell apoptosis. Meanwhile muscle fiber atrophy with mitochondria vacuolation was found in aging rat by electron microscope.
Subject(s)
Aging , Gene Expression Profiling , Muscle, Skeletal/metabolism , Animals , Muscle, Skeletal/growth & development , Oligonucleotide Array Sequence Analysis/methods , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Sprague-DawleyABSTRACT
Human Prostate Specific Membrane Antigen(PSMA) cDNA was amplified using total RNA extracted from prostate carcinoma tissue by RT-PCR. The cDNA fragment of extracellular domain of PSMA(edPSMA) gene was amplified by PCR and cloned into expression vector pMAL-c2x. Sequence analysis of both PSMA and edPSMA revealed identity to the GenBank reported. The edPSMA was expressed in E. coli as part of a fusion protein with MBP as the induction of IPTG. Western blot analysis showed the recombinant protein could react with PSMA monocloned antibodies 4G5. MBP-edPSMA fusion protein were purified by amylose resin affinity chromatography and showed to be homogeneity in SDS-PAGE(120 kD). BALB/C mice were immunized with the purified protein for the preparation of polyclonal antibody. The polyclonal antibody, which had a title of 1:12,800, were indicated the specificity to prostate tissue.
