The hsDC2, an unknown gene, was located on chromosome 4q25. Its genetic protein product contains 149 amino acids with the molecular weight of 16.8 kDa approximately. Predicted by bioinformatics, hsDC2 might be a transmembrane protein with three transmembrane helices on the endoplasmic reticulum. Based on the results of reverse transcription-polymerase chain reaction, it revealed that hsDC2 was expressed in many tissues. It was showed that there were more mRNA expression in the cancer tissue. Using real-time quantitative PCR to analyze cancer cell lines, we found that hsDC2 might be related to differentiation status of cells. Among the well-differentiated cells such as nasopharyngeal carcinoma cell line NPC TW01, hepatoma cell line Hep3B and temperature-induced differentiated human fetal osteoblast cell line (hFOB), there were more hsDC2 mRNA expression. We also obtained some useful information from bioinformatics databases. To further elucidate the biological functions of the gene, glutathion S-transferase-hsDC2 fusion protein was used to generate anti-hsDC2 polyclonal antibody. However, we still need further research to clarify the biological function of hsDC2.

The hsFLJ22386 gene, an unknown gene, was located on chromosome 16p13.3. Its protein product contains 287 amino acids with the molecular weight of 32.2 kDa approximately. Predicted by bioinformatics, hsFLJ22386 might be a protein containing a leucine zipper domain. Based on the results of reverse transcriptase polymerase chain reaction, it revealed that FLJ22386 was expressed in several nervous system tissues and several organ tissues (liver, spleen, small intestine and kidney). In human cancer cell lines, the RT-PCR results showed that FLJ22386 was expressed in brain tumor cell lines (T98G, U87MG, U251, GBM8401), nasopharyngeal epithelial cell line (NNE-3)and carcinoma cell lines (NPCTW01, NPCTW04), hepatoma cell lines (J5, Hep3B, SK-Hep-1) and lymphoma cell lines (RPMI, P3HR1, Raji, U937). Human FLJ22386 coding sequence was inserted into pEGFP-C2 plasmid, and the tag-fused gene was transfected into NIH3T3 cells to see if it has the ability to promote cell proliferation. To further investigate the protein level expression and biological functions of the gene, glutathione S-transferase-hsFLJ22386 fusion protein was expressed and used to generate anti-FLJ22386 polyclonal antibody. According to the results of RT-PCR and anchorage dependent growth assay, it is presumed that FLJ22386 may play a role in cell proliferation.

Cell walls are a distinct feature of plants and their chemical constituents, cellulose, hemicelluloses and lignin, are economically valuable. Plant fibres rich in cellulose, which mainly resides in their cell wall, are traditionally used in making paper and textiles. The changing global economic situation and environmental concerns have imparted necessity for renewable, but at the same time value added cellulosic materials. The Department of Wood Biotechnology, KTH together with its collaborators, have established EST libraries and performed transcript profiling during wood development in poplar, a tree considered as a model for wood development. The majority of the genes upregulated during cellulose biosynthesis encode proteins with known or predictable functions, such as carbohydrate active enzymes (CAzymes). However, some of them encode proteins with unknown functions. Characterization of these genes will potentially give additional opportunities to modify fibre properties. This thesis describes the discovery and characterization of a highly upregulated gene with a previously unknown function in poplar xylem, here denoted PttMAP20. Following its early discovery by mRNA profiling, the characterization was initiated with a thorough bioinformatic analysis, and the knowledge obtained was used to devise techniques for further functional analysis. Specific antibodies were raised, affinity purified and characterized. The antibodies were used as a tool for screening recombinant expression in E. coli and for the cellular localization of the protein in plant tissues, visualized with confocal and transmission electron microscopy. A purification protocol was developed for the expressed protein, followed by biochemical characterization. Appropriate model systems were used in both in vivo and in vitro studies. Fluorescently labelled protein transiently expressed in tobacco leaves was used for localization studies and the same system was used to characterize the molecular properties of the protein. Phenotypes arising from overexpressing the PttMAP20 gene were traced in the model plant Arabidopsis. All the results obtained so far indicate that PttMAP20 is a novel microtubule associated protein that binds to a cellulose biosynthesis inhibitor, DCB (2,6-dichlorobenzonitrile) and is required during cellulose biosynthesis in secondary cell walls. / QC 20100906