Méjean V, Claverys J P, Vasseghi H, Sicard A M
Gene. 1981 Nov;15(2-3):289-93. doi: 10.1016/0378-1119(81)90139-6.
A method for the rapid cloning of specific Streptococcus pneumoniae DNA fragments depends on the integration by homologous recombination into the bacterial chromosome of a plasmid which carries an insert of S. pneumoniae DNA, but which cannot be autonomously maintained in S. pneumoniae. Selection for plasmid integration employs aminopterin or erythromycin resistance. Host sequences adjacent to the site of insertion are easily cloned by enzymatic excision and recircularization of the plasmid, followed by propagation in Escherichia coli. This is particularly useful for repeated cloning of a given fragment that carries various mutations.
一种快速克隆特定肺炎链球菌DNA片段的方法，依赖于通过同源重组将携带肺炎链球菌DNA插入片段但不能在肺炎链球菌中自主维持的质粒整合到细菌染色体中。通过氨甲蝶呤或红霉素抗性来选择质粒整合。通过对质粒进行酶切切除和重新环化，然后在大肠杆菌中繁殖，可轻松克隆与插入位点相邻的宿主序列。这对于重复克隆携带各种突变的给定片段特别有用。
