pACYCDuet-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p15A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。
pACYCDuet-1 is designed for the coexpression of two target genes. The vector contains two multiple cloning sites (MCS), each of which is preceded by a T7 promoter/lac operator and ribosome binding site (rbs). The vector also carries the P15A replicon, lacI gene and chloramphenicol resistance gene. This vector can be used in combination with pETDuet™-1 in an appropriate host strain for the coexpression of up to 4 target genes. Genes inserted into MCS1 can be sequenced using the ACYCDuetUP1 Primer and DuetDOWN1 Primer . Genes inserted into MCS2 can be sequenced using the DuetUP2 Primer and T7 Terminator Primer .