RESUMEN
Ubiquitous chromatin opening element ( UCOE ) , composed of the promoters of human housekeeping genes, prevents transgene from silencing and produces consistent, stable and high-level gene expression irrespec-tive of the chromosomal integration site. The research studied the influence of different UCOE element parts on antibody expression in CHO cells. UCOE 1. 5 kb from chromobox homolog 3 ( CBX3 ) , UCOE 2. 5 kb from the heterogeneous ribonucleoprotein A2/B1 ( HNRPA2 B1 ) and the whole UCOE 4. 0 kb were inserted into the anti-body light and heavy chain vectors, respectively, and transfected into CHO cells using antibiotics-Zeocin and Blasticidin for pressure selection. Four groups of monoclonal cells were harvested and antibody expression of each group was detected. The monoclonal cells with UCOE 1. 5 kb and UCOE 2. 5 kb increased 1. 5 to 2-fold in the level of antibody expression, wheareas, monoclonal cells with UCOE 4. 0 kb increased 3 to 4-fold. The enhance-ment of two housekeeping promoter genes on antibody expression could stack up.
RESUMEN
Multilocus sequence typing MLST with high solution sensitivity and specificity is widely used to study the population genetic structure of pathogen by amplification and sequencing of the housekeeping genes. MLST also provides more evidence and plays an important role in parasite research. This paper reviews the principle and method of MLST and its applica-tion on population genetic structure analysis of parasites.
RESUMEN
Objective:To investigate the effect of pcDNA3.1 vector on house keeping gene expression and set guide to further study work in using pcDNA3.1. Methods:5 cell lines were transfected with pcDNA3.1,48 h post transfection total RNA was extracted and applied for real time PCR to detect the RNA level of GAPDH、 ?-actin、18sRNA difference.Meanwhile,extract total protein and analyze the GAPDH and?-actin expression by Western blot.Results:RNA and protein levels all show of a decrease in 5 cell lines after transfection,with the most obviously ?-actin.Conclusion:pcDNA3.1 might have non specifinhibition effect to cell,when using pcDNA3. 1 for gene function research,it is not recommend to use ?- actin as reference gene.