Expression and Localization of BALB/c Mouse T Cell Immunoglobulin and Mucin Domain Containing Molecules-2 in L929 Cell Line and Facilitate the Ferritin Endocytosis
|School||Huazhong University of Science and Technology|
|Course||Clinical Laboratory Science|
|Keywords||Tim-2 gene BALB / c mice L929 cells Eukaryotic expression Ferritin Kunming mice Single nucleotide polymorphisms|
First portion of mice Tim - 2 gene in the expression of L929 cells to promote endocytosis of the ferritin Objective: To obtain the T cells of BALB / c mouse immunoglobulin domain sticky protein domain protein - ( 2T cell Immunoglobulin and Mucin domain containing the molecules -2 Tim-2) gene construct eukaryotic expression vector expressed in L929 cells and study their function . Methods : RT-PCR for BALB / c mice Tim -2 gene digested after sequencing the TA cloning , inserted into the eukaryotic expression vector pEGFP-N1 liposomal transfection of L929 cells , fluorescence microscopy exogenous gene expression , and then added the role of ferritin transfected cells . Results: exogenous gene was highly expressed in L929 cells , fluorescence microscopy results show that the exogenous gene is located on the cell membrane and promote cell ferritin within swallow . Conclusion : Tim -2 gene expression in eukaryotic cells , positioned on the membrane , and to promote cell ferritin inner swallow . The second part of Kunming mice Tim-2 gene single nucleotide variations Objective: To study the T cell immunoglobulin domain sticky protein domain protein (T cell immunoglobulin-and mucin-domain-containing molecules, Tim) gene family members one - Tim-2 gene single nucleotide polymorphisms in Kunming mice prompted Kunming mice in oncology and immunology tests internationally accepted other strains of mice ( BALB / c mice ) different biological characteristics . Method: five different sources Kunming mice liver tissue , total RNA was extracted , reverse transcription to obtain cDNA, Tim-2 gene was amplified by PCR using specific primers , cloned and sequenced , and then compare the sequencing map . Results: The sequencing results in Genebank Tim-2 sequence , 451 (TCC → GCC : serine → alanine ) , 502, 504 (CCA → TCT : proline → serine ) , 586,587 ( GAT → TCT : aspartyl acid → serine ) . Other amino acid changes in the variable sites . Conclusion: Tim -2 protein encoded by the gene sequences of other strains of mice ( BALB / c mice ) prompted Kunming mice with the international common differences in the immunological test in Kunming mice should noted that because of this difference caused by the impact on the experimental results .