摘要
The potted Leymus chinensis seedlings were treated with saline-alkali solution of six different(from Ⅰ to Ⅵ) concentrations. The results demonstrate that the betaine content and Betaine-aldehyde dehydrogenase(BADH: EC 1.2.1.8) activities have a direct relation with increased stressing time in the same treatment; both exhibit a single peak with increasing the concentration of saline-alkali solution, and number V shows the highest value. The BADH gene of Leyrnus chinensis was cloned by RT-PCR and RACE technology and was designated as LcBADH. The cDNA sequence of LcBADH was 1774bp including the open reading frame(ORF) of 1521bp(coding 506 amino acids). The vector of prokaryotic expression was constructed by inserting the LcBADH gene fragmcnt into pET30a(+) and transformed into E. coli BL21(DE3). The result of SDS-PAGE shows that the idio-protein with a molecular mass of 56.78 kDa was effectively expressed in the recombinant bacteria induced by isopropyl fl-D-thiogalactoside(IPTG).
The potted Leymus chinensis seedlings were treated with saline-alkali solution of six different(from Ⅰ to Ⅵ) concentrations. The results demonstrate that the betaine content and Betaine-aldehyde dehydrogenase(BADH: EC 1.2.1.8) activities have a direct relation with increased stressing time in the same treatment; both exhibit a single peak with increasing the concentration of saline-alkali solution, and number V shows the highest value. The BADH gene of Leyrnus chinensis was cloned by RT-PCR and RACE technology and was designated as LcBADH. The cDNA sequence of LcBADH was 1774bp including the open reading frame(ORF) of 1521bp(coding 506 amino acids). The vector of prokaryotic expression was constructed by inserting the LcBADH gene fragmcnt into pET30a(+) and transformed into E. coli BL21(DE3). The result of SDS-PAGE shows that the idio-protein with a molecular mass of 56.78 kDa was effectively expressed in the recombinant bacteria induced by isopropyl fl-D-thiogalactoside(IPTG).
基金
Supported by the National Natural Science Foundation of China(Nos30590382 and 30570273)