Identification and nucleotide sequence of Rhizobium meliloti insertion sequence ISRm3: similarity between the putative transposase encoded by ISRm3 and those encoded by Staphylococcus aureus IS256 and Thiobacillus ferrooxidans IST2
Identification and nucleotide sequence of Rhizobium meliloti insertion sequence ISRm3: similarity between the putative transposase encoded by ISRm3 and those encoded by Staphylococcus aureus IS256 and Thiobacillus ferrooxidans IST2
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苜蓿根瘤菌插入序列ISRm3的鉴定和核苷酸序列：ISRm3编码的推定转座酶与金黄色葡萄球菌IS256和氧化亚铁硫杆菌IST2编码的转座酶之间的相似性
DOI：
10.1128/jb.173.8.2530-2538.1991
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发表时间：
1991
影响因子：
3.2
通讯作者：
Serge Laberge
中科院分区：
文献类型：
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作者：
Roger WHEATCROFTl;Serge Laberge
用~(32)P标记的pTA2对细胞总DNA进行Southern杂交，在不同的紫花苜蓿根瘤菌中同时发现了插入序列ISRm3。该质粒是IZ450菌株所特有的，并且碰巧含有四个拷贝的ISRm3。通过使用内部EcoRI片段作为特异性探针(PRWRm31)，随后在来自世界不同地理位置的90%以上的苜蓿根瘤菌菌株中检测到与ISRm3的同源性。在液体培养条件下，稳定的非致死性ISRM3转位的频率估计为每代细胞4×10(-5)。紫花苜蓿102F70的ISRm3基因全长1,298个碱基，末端有30个碱基的反向重复序列，完全匹配。对两个菌株的6个ISRm3拷贝的分析表明，不同数量的碱基对(通常是8个或9个)重复，并在插入部位附近形成直接重复。在一条DNA链上，ISRm3含有一个长度为93%的开放阅读框。将推测的蛋白与EMBL和GenBank数据库中的序列进行比较，发现苜蓿根瘤菌的ISRM3、金黄色葡萄球菌的IS256和铁硫杆菌的IST2的转座酶具有显著的相似性。这些插入序列似乎是不同类别的远亲成员。
The insertion sequence ISRm3 was discovered simultaneously in different Rhizobium meliloti strains by probing Southern blots of total cellular DNA with 32P-labeled pTA2. This plasmid is indigenous to strain IZ450 and fortuitously contained four copies of ISRm3. By using an internal EcoRI fragment as a specific probe (pRWRm31), homology to ISRm3 was subsequently detected in over 90% of R. meliloti strains tested from different geographical locations around the world. The frequency of stable nonlethal ISRm3 transpositions was estimated to be 4 x 10(-5) per generation per cell in strain SU47 when grown in liquid culture. The entire nucleotide sequence of ISRm3 in R. meliloti 102F70 is 1,298 bp and has 30-bp terminal inverted repeats which are perfectly matched. Analysis of six copies of ISRm3 in two strains showed that a variable number of base pairs (usually eight or nine) were duplicated and formed direct repeats adjacent to the site of insertion. On one DNA strand, ISRm3 contains an open reading frame spanning 93% of its length. Comparison of the putative protein encoded with sequences derived from the EMBL and GenBank databases showed significant similarity between the putative transposases of ISRm3 from R. meliloti, IS256 from Staphylococcus aureus, and IST2 from Thiobacillus ferroxidans. These insertion sequences appear to be distantly related members of a distinct class.
M.Maejima；M.Inoue：质粒。
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