邻苯二酚1,2-双加氧酶(C12O)基因的定位、克隆和表达

LOCATION,CLONING AND EXPRESSION OF CATECHOL 1,2 DIOXYGENASE (C12O) GENE

射性同位素标记ｔｆｄＣ基因片段作探针，Ｓｏｕｔｈｅｒｎｂｌｏｔ杂交定位Ｌ１菌株的邻苯二酚１，２双加氧酶基因位于ＰｓｔⅠ的Ｉ片段和ＢａｍＨⅠ的Ｍ和Ｎ片段．低熔点琼脂糖回收ＰｓｔⅠ的Ｉ段，直接克隆至表达载体ｐＫＴ２３０上，获得重组子ｐＫＴ２３０ｐ．重组子转化不具开环酶活性的甲胺磷降解菌Ｐ２。

The tfd C fragment was labelled by radioisotope as a probe.The C12O gene was identified to be on the Pst Ⅰfragment Ⅰand Bam H Ⅰ fragments M and N of pL1 by Southern blot hybridization. Pst Ⅰ fragment was reclaimed by agarose LMP and cloned on the pKT230.The recombinant was transformed into P 2 strain,which can degrade methamidophos and has no ring cleavage enzyme activities.The transformant harbored hybrid plasmid and showed high level enzyme activities.