Construction of decorin expression vector and its expression in CHO cells / 吉林大学学报(医学版)

Objective To construct a eukaryotic expression vector of decorin(DCN),and observe its expression in CHO cells,in order to provide a basis for further study on the anti-tumor effect of DCN. Methods DCN cDNA was amplified by PCR.The human full-length DCN cDNA ligated into pBluescript was used as template.The fragment was ligated to the expression vector pCDNA3 previously digested with XbaⅠ and EcoR Ⅰ.The ligation mixture was transformed into competent E.coli JM109 cells.Transformants containing inserts were confirmed by restrictive digestion and DNA sequencing.The expression vector was transfected into CHO cells using lipofectamine,and transfected cells were cultivated in DMEM containing G418 (800 mg?L-1) for about 2 months.Immunohistochemistry method was used to detect the expression of DCN protein in stably transfected cells.Results The PCR product was about 1 000 bp.The recombinant expression vector was identified by restrictive digestion and DNA sequencing. DCN protein was detectable in stablely transfected cells.Conclusion The recombinant eukaryotic expression vector pCDNA-DEC is constructed successfully and stablely transfected CHO cells are established.

Construction and identification of human decorin gene recombinant eukaryotic expressing vector / 吉林大学学报(医学版)

Objective To construct a recombinant eukaryotic expressing vector pcDNA-dec and provide a basis for further study on the bioactivity of decorin(DCN).Methods DCN cDNA was amplified by using polymerase chain reaction(PCR).Product of PCR and expressing vector were digested by restriction endonucleases,then ligated and transformed into JM109 bacteria.Results The specific DNA fragment was obtained by PCR as supposed.Product of PCR and expressing vector were digested by restriction endonucleases,then ligated to establish the recombinant eukaryotic expression vector pcDNA-dec.It was confirmed that DCN cDNA was inserted into the eukaryotic expression vector correctly by using digestion identification and sequencing.Conclusion The recombinant eukaryotic expression vector pcDNAdec of human DCN is successfully constructed.

Expressions of decorin mRNA and protein in colorectal carcinoma / 吉林大学学报(医学版)

0.05).DCN protein didn′t express in both 20 cases of relative normal tissue and(30 cases) of colorectal carcinoma.Conclusion DCN mRNA can express in both colorectal carcinoma and relative normal tissue.