pRSFDuet-1质粒是一种原核表达载体, N端含有一个6×His标签,C端含有一个S标签。该质粒含有多个常用的酶切位点,便于不同基因克隆。表达由宿主细胞提供的T7 RNA聚合酶诱导,目的基因被克隆到质粒载体上,受噬菌体强转录及翻译信号控制。
该质粒还携带有RSF复制因子、lacI基因和氯霉素抗性基因。可以使这个pETDuet-1载体在一个适当的宿主菌株的coexpression 4目标基因插入MCS1的基因可以使用ACYCDuetUP1引物和DuetDOWN1引物进行测序。插入MCS2的基因可以用DuetUP2引物和T7进行排序。
pRSFDuet-1 is designed for the coexpression of two target ORFs. The vector contains two multiple cloning sites (MCS), each of which is preceded by a T7lac promoter and ribosome binding site (rbs). The vector also carries the RSF1030-derived RSF replicon, lacI gene and kanamycin resistance gene (KnR). This vector can be used in combination with pACYCDuet™-1 , pCDFDuet™-1 , and pETDuet™-1 in an appropriate host strain for the coexpression of 4 to 8 target proteins. ORFs inserted into MCS1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer . ORFs inserted into MCS2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer . Unique sites are shown on the circle map.