Three transfection reagents, Lipofectamine® 2000, TransIT-PRO® and linear 25 kDa polyethylenimine were evaluated for transient expression of enhanced green fluorescent protein in Chinese hamster ovary ce...Three transfection reagents, Lipofectamine® 2000, TransIT-PRO® and linear 25 kDa polyethylenimine were evaluated for transient expression of enhanced green fluorescent protein in Chinese hamster ovary cells. TransIT-PRO® was found to be more efficient under the examined conditions, but comes at an increased cost compared to the widely used PEI.展开更多
Aim: To study the characteristics and possible retention function of specific sequence in the 5'-end of rat testis GABAA receptor β 3t variant. Methods: Rat testis GABAA receptor β 3t variant cDNA was cloned and...Aim: To study the characteristics and possible retention function of specific sequence in the 5'-end of rat testis GABAA receptor β 3t variant. Methods: Rat testis GABAA receptor β 3t variant cDNA was cloned and inserted into two eukaryotic expression vectors of pEGFP-Nl and pEGFP-Cl respectively, which have EGFP reporter gene. The recombinant plasmids were transfected into Chinese hamster ovary (CHO) cells with the calcium phosphate co-precipitation method. Fluorescence microscope and laser confocal microscope were used to analyze the transfected cells. ConA-Texas-Red was used to label cell endoplasmic reticulum (ER) and study the located area of rat testis β 3t variant in the CHO cells. Results: When the rat testis β 3t variant express in CHO cells, there were two expression pattern, the even and the concentrated expression pattern. Although rat brain β 3 can reach cell membrane, the rat testis β 3t variant can't target to the cell membrane. It may be retained in the ER of CHO cells. Conclusion: There may exist an ER retention signal in the 25 specific amino acid sequence in the N terminal of rat testis β 3t variant.展开更多
Objective : To study the antitoxic role of vesicular monoamine transporter 2 (VMAT2) in transgenic Chinese Hamster ovary (CHO) cell. Methods :With the technology of trans-gene from PC 12 to CHO, MTT reduction assay wa...Objective : To study the antitoxic role of vesicular monoamine transporter 2 (VMAT2) in transgenic Chinese Hamster ovary (CHO) cell. Methods :With the technology of trans-gene from PC 12 to CHO, MTT reduction assay was used to detect MPP+ toxic effect on wild type CHO (wtCHO) and transgenic CHO. Meanwhile, the role of reserpine was also observed in MPP+ toxic effects. Results :The sensitivity of transgenic CHO to MPP+ was much less than that of wtCHO with 0. 5 mmol/L MPP+. Transgenic CHO had the same sensitivity as wtCHO if rotenone was given. WtCHO, by given reserpine alone, didn't change its sensitivity to MPP+. Conclusions :VMAT2 has protective effect on transgenic CHO by transporting MPP+ to vesicles.展开更多
文摘Three transfection reagents, Lipofectamine® 2000, TransIT-PRO® and linear 25 kDa polyethylenimine were evaluated for transient expression of enhanced green fluorescent protein in Chinese hamster ovary cells. TransIT-PRO® was found to be more efficient under the examined conditions, but comes at an increased cost compared to the widely used PEI.
文摘Aim: To study the characteristics and possible retention function of specific sequence in the 5'-end of rat testis GABAA receptor β 3t variant. Methods: Rat testis GABAA receptor β 3t variant cDNA was cloned and inserted into two eukaryotic expression vectors of pEGFP-Nl and pEGFP-Cl respectively, which have EGFP reporter gene. The recombinant plasmids were transfected into Chinese hamster ovary (CHO) cells with the calcium phosphate co-precipitation method. Fluorescence microscope and laser confocal microscope were used to analyze the transfected cells. ConA-Texas-Red was used to label cell endoplasmic reticulum (ER) and study the located area of rat testis β 3t variant in the CHO cells. Results: When the rat testis β 3t variant express in CHO cells, there were two expression pattern, the even and the concentrated expression pattern. Although rat brain β 3 can reach cell membrane, the rat testis β 3t variant can't target to the cell membrane. It may be retained in the ER of CHO cells. Conclusion: There may exist an ER retention signal in the 25 specific amino acid sequence in the N terminal of rat testis β 3t variant.
基金Supported by grant from Innovation Foundation of Nanjing Medical University(MC9901)
文摘Objective : To study the antitoxic role of vesicular monoamine transporter 2 (VMAT2) in transgenic Chinese Hamster ovary (CHO) cell. Methods :With the technology of trans-gene from PC 12 to CHO, MTT reduction assay was used to detect MPP+ toxic effect on wild type CHO (wtCHO) and transgenic CHO. Meanwhile, the role of reserpine was also observed in MPP+ toxic effects. Results :The sensitivity of transgenic CHO to MPP+ was much less than that of wtCHO with 0. 5 mmol/L MPP+. Transgenic CHO had the same sensitivity as wtCHO if rotenone was given. WtCHO, by given reserpine alone, didn't change its sensitivity to MPP+. Conclusions :VMAT2 has protective effect on transgenic CHO by transporting MPP+ to vesicles.
