摘要
从一株氧化亚铁硫杆菌(Thiobacillus ferrooxidans)中分离到一个分子量为4.5×10~6Md 的质粒 pTf—52,用 Pst Ⅰ、BglⅡ、Kpn Ⅰ酶切此质粒,进行限制性酶切分析,作出了 pTf—52的限制图谱;并将 pTf—52插入到 pBR325的 Pst Ⅰ位点,体外重组,转化 E.coli HB101,构建成一个能够在 E.coli 中进行复制的嵌合质柱 pSDF—1(Tc^rCm^rAp^s),分子量为8.2×10~6Md。经限制性酶切分析,确证 pSDF—1是由 pBR325和 pTf—52重组成的,并确定了这两种质粒在 pSDF—1中的连接方向。
A 4.5×10~6 daltons plasmid,pTf—52,was isolated from a T.ferrooxi- dans strain.Physical mapping of pTf—52 was made by restriction analysis using Pst Ⅰ,Bgl Ⅱ and Kpn Ⅰ.A chimeric plasmid pSDF—1(Tc^r Cm^r Ap^s) which had the M.W of 8.2×10~6 daltons was constructed by insertion of pTf—52 into the PSt Ⅰ site of the plasmid pBR325,followed by transfor- mation into E.coli HB101.By restriction analysis,pSDF—1 was further confirmed and insertion direction of pTf—52 in pBR325 was also determined.
出处
《山东大学学报（自然科学版）》
CSCD
1990年第3期389-394,共6页
Journal of Shandong University(Natural Science Edition)
关键词
硫杆菌
氧化亚铁
限制图谱
脱硫
T.ferrooxidans
restriction mapping
chimeric plasmid
