An expression vector pACⅢ containing the chimeric gene HAV- VP3P1 and HCV-C gene has been constructed and transferred to C. reinhardtii by the biolistic method. The trans-formants have been identified by PCR, Souther...An expression vector pACⅢ containing the chimeric gene HAV- VP3P1 and HCV-C gene has been constructed and transferred to C. reinhardtii by the biolistic method. The trans-formants have been identified by PCR, Southern-blotting, Northern-blotting and Western-blotting assays after selecting on resistant medium and incubating in the dark. The results展开更多
The aim of this study was to investigate the mechanism of cadmium (Cd) adsorbed by microalgae Chlamydomonas reinhardtii (C.reinhardtii). The kinetic and adsorption isotherm of the process could be well described by ma...The aim of this study was to investigate the mechanism of cadmium (Cd) adsorbed by microalgae Chlamydomonas reinhardtii (C.reinhardtii). The kinetic and adsorption isotherm of the process could be well described by mathematical models. Chemical modification experiments and Fourier transform infrared spectra indicated that carboxyl and amine groups were the important functional groups for adsorption of Cd. The maximum contribution of physical adsorption in the overall adsorption process was evaluated as 5.5%. These results indicated that chemisorption was the dominating mechanism of Cd biosorption by C.reinhardtii.展开更多
Tumor necrosis factor-related apop-tosis-inducing ligand (TRAIL) induces selectively apoptosis in various tumor cells and virus-infected cells, but rarely in normal cells. A chloroplast expres-sion vector, p64TRAIL, c...Tumor necrosis factor-related apop-tosis-inducing ligand (TRAIL) induces selectively apoptosis in various tumor cells and virus-infected cells, but rarely in normal cells. A chloroplast expres-sion vector, p64TRAIL, containing the cDNA coding for the soluble TRAIL (sTRAIL), was constructed with clpP-trnL-petB-chlL-rpl23-rpl2 as Chlamydomonas reinhardtii plastid homologous recombinant frag-ments and spectinomycin-resistant aadA gene as a select marker. The plasmid p64TRAIL was trans-ferred into the chloroplast genome of C. reinhardtii by the biolistic method. Three independently trans-formed lines were obtained by 100 mg/L spectino-mycin selection. PCR amplification, Southern blot analysis of the sTRAIL coding region DNA and culti-vation cells in the dark all showed that the exogenous DNA had been integrated into chloroplast genome of C. reinhardtii. Western blot analysis showed that human soluble TRAIL was expressed in C. reinhardtii chloroplast. The densitometric analysis of Western blot indicated that the expressed human sTRAIL protein in the chloroplasts of C. reinhardtii accounted for about 0.43%―0.67% of the total soluble proteins. These experimental results demonstrated the possi-bility of using transgenic chloroplasts of green alga as bioreactors for production of biopharmaceuticals.展开更多
文摘An expression vector pACⅢ containing the chimeric gene HAV- VP3P1 and HCV-C gene has been constructed and transferred to C. reinhardtii by the biolistic method. The trans-formants have been identified by PCR, Southern-blotting, Northern-blotting and Western-blotting assays after selecting on resistant medium and incubating in the dark. The results
基金supported by Beijing Elitist Foundation (Grant no. 2005IA05016012) Beijing New Century Talent Foundation (Grant no. 011 005400601)
文摘The aim of this study was to investigate the mechanism of cadmium (Cd) adsorbed by microalgae Chlamydomonas reinhardtii (C.reinhardtii). The kinetic and adsorption isotherm of the process could be well described by mathematical models. Chemical modification experiments and Fourier transform infrared spectra indicated that carboxyl and amine groups were the important functional groups for adsorption of Cd. The maximum contribution of physical adsorption in the overall adsorption process was evaluated as 5.5%. These results indicated that chemisorption was the dominating mechanism of Cd biosorption by C.reinhardtii.
基金supported by the National Natural Science Foundation of China(Grant No.30571189)a grant from Tianjin Municipal Science and Technology Commission(Grant No.033605111)to Yong Wang.
文摘Tumor necrosis factor-related apop-tosis-inducing ligand (TRAIL) induces selectively apoptosis in various tumor cells and virus-infected cells, but rarely in normal cells. A chloroplast expres-sion vector, p64TRAIL, containing the cDNA coding for the soluble TRAIL (sTRAIL), was constructed with clpP-trnL-petB-chlL-rpl23-rpl2 as Chlamydomonas reinhardtii plastid homologous recombinant frag-ments and spectinomycin-resistant aadA gene as a select marker. The plasmid p64TRAIL was trans-ferred into the chloroplast genome of C. reinhardtii by the biolistic method. Three independently trans-formed lines were obtained by 100 mg/L spectino-mycin selection. PCR amplification, Southern blot analysis of the sTRAIL coding region DNA and culti-vation cells in the dark all showed that the exogenous DNA had been integrated into chloroplast genome of C. reinhardtii. Western blot analysis showed that human soluble TRAIL was expressed in C. reinhardtii chloroplast. The densitometric analysis of Western blot indicated that the expressed human sTRAIL protein in the chloroplasts of C. reinhardtii accounted for about 0.43%―0.67% of the total soluble proteins. These experimental results demonstrated the possi-bility of using transgenic chloroplasts of green alga as bioreactors for production of biopharmaceuticals.
