《Table 1 Primers for construction of plasmids》

《Table 1 Primers for construction of plasmids》   提示:宽带有限、当前游客访问压缩模式
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《蓝藻红色荧光蛋白All1280 GAF2在E. coli BL21(DE3)中的表达及其突变体构建(英文)》


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Primers were designed based on the nucleotide sequence of all1280 gaf 2(Table 1).Using the genomic DNA of Anabaena sp.PCC 7120 as a template,all1280 gaf2 was amplified with primers P1-P2.The all1280 gaf2 was then inserted into pET-30a(+)using Bam HⅠand SalⅠenzyme sites,with pET-all1280 gaf2 thus obtained.For site-directed mutagenesis,P3 was designed as the conserved C53 replaced with A53.Using the site-directed mutagenesis kit,all1280 gaf2(C53A)was amplified with pET-all1280 gaf2 as the template,P3 and P4 as the primers.The all1280 gaf2(C53A)was then inserted into pET-30a(+)using Bam HⅠand SalⅠ,with pET-all1280 gaf 2(C53A)thus constructed.Nucleotide sequencing identified that all1280 gaf 2 and all1280 gaf2(C53A)fragments were successfully cloned into the pET-30a(+)vector.