摘要
丝状蓝藻plectonemaboryanum的pPb1经HpaⅡ酶酶切与AccⅠ酶酶切的pUC13构成杂合质粒,杂合质粒克隆到E.coliJM109、频率为3.4%.转化进E.ColiJM109的杂合质粒,用EcoRⅠ和HpaⅠ酶切后得到证实.杂合质粒转化到新制备的P.boryanum原生质球或丝状体后,再生的藻体表现出高于原来两倍以上的Ap抗性.利用Ap敏感的E.coli平板培养,证实了转化入藻体的质粒对Ap具有高抗性.然而,获得的Ap高抗株在后续培养中不能继代维持.
Hybrid-plasmid from Hpa Ⅱ-digested ppb_1and AccⅠ-cleaved pUC13 was cloned into E. coli JM109. The purified hybrid-plasmid was transformed backwardly into freshly prepared spheroplast or filament from P. boryanum. The transformed algal colonies displayed anti-ampicillin activity in a level up to two times higher concentration of Ap had been confirmed in plate culture with an AP sensitific E. coli strain. Nevertheless,the acquired Ap ̄r can not withstand this level of AP on longer succesive culture.
出处
《厦门大学学报(自然科学版)》
CAS
CSCD
北大核心
1994年第1期105-110,共6页
Journal of Xiamen University:Natural Science
基金
国家自然科学基金
关键词
蓝藻
大肠杆菌
杂质粒
构建
克隆
Plectonema boryanum,pUC_13-pPb_1, Transformation