| Summary: | 碩士 === 國立暨南國際大學 === 生物醫學科技研究所 === 95 === Cancer is one of the human diseases with high prevalence in the world, among them, lung cancer is the leading cause of cancer death around the world as well as in Taiwan. The previous evidence proved that cancer has a relationship with multi-gene mutation. In our previous studies, using lung cancer tissue and its’ adjacent non-tumor part with microarray, we found some genes with different expression level in both part. SLR417 is one of those genes which is down-regulated in tumor part as compared to adjacent normal part. Combined the microarray and Q-PCR analysis data and some SLR417-related informations, we hypothesize that SLR417 maybe a novel tumor suppressor gene. To study this novel gene, we used yeast two hybrid assay which is one method to study protein-protein interaction in vivo to find those meaningful interaction protein. We constructed the full-length human SLR417 gene to fuse in-frame with the GAL4 DNA binding domain as bait, a commercial human testis cDNA library as prey. Our results show that there are 717 colonies selectively grew on QDO plates (Ade.His.Leu.Trp nutritionally deficient plates) and 54 colonies among those could also activate - and -galactosidase. Subsequently, we sequencing and BLAST analysized those colonies. Now 6 suspected SLR417 interaction proteins [accession number of these genes as following: NM_014939 (KIAA1012)、NM_006432 (NPC2)、AB209134 (Jab1)、BC131805 (coiled-coil domain containing 46)、BC019827 (DNAJB1)、NM_016299 (HSPA14) ] were identified and under further examined.Among them, Jab1 has been confirmed to interact with SLR417 in vitro cell culture system by immunoprecipitation analysis.
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