A new Thermus sp class-IIS enzyme sub-family:: isolation of a 'twin' endonuclease TspDTI with a novel specificity 5′-ATGAA(N11/9)-3′, related to TspGWI, TaqII and Tth111II -: art. no. e74
A new Thermus sp class-IIS enzyme sub-family:: isolation of a 'twin' endonuclease TspDTI with a novel specificity 5′-ATGAA(N11/9)-3′, related to TspGWI, TaqII and Tth111II -: art. no. e74
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DOI：
10.1093/nar/gng074
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发表时间：
2003-07-15
影响因子：
14.9
通讯作者：
Harasimowicz-Slowinska, RI
中科院分区：
文献类型：
--
作者：
Skowron, PM;Majewski, J;Harasimowicz-Slowinska, RI
从热菌（Thermus sp. DT）中分离到具有5‘-ATGAA(N-11/9)-3’特异性的TspDTI限制性内切酶。正如我们之前报道的那样，TspDTI似乎是来自Thermus sp. GW的限制性内切酶TspGWI的“双胞胎”。TspGWI与TspDTI是从同一位置分离出来的，它识别一个相关序列5'-ACGGA(N-11/9)-3'，并且具有保守的切割位置。这两种酶类似于来自热菌属的另外两种iis类内切酶：TaqII和Tth111II。TspGWI色氨酸n端氨基酸序列与TaqII序列的相似性为88.9-100%。所有四种酶均纯化至均匀性;它们的多肽大小（114.5-122 kDa）使它们成为迄今为止已知的最大的ii类限制性内切酶。本文提出了一个具有共同起源的iis类限制性内切酶亚家族的存在。
The TspDTI restriction endonuclease, which shows a novel recognition specificity 5'-ATGAA(N-11/9)-3', was isolated from Thermus sp. DT. TspDTI appears to be a 'twin' of restriction endonuclease TspGWI from Thermus sp. GW, as we have previously reported. TspGWI was isolated from the same location as TspDTI, it recognizes a related sequence 5'-ACGGA(N-11/9)-3' and has conserved cleavage positions. Both enzymes resemble two other class-IIS endonucleases from Thermus sp.: TaqII and Tth111II. N-terminal amino acid sequences of TspGWI tryptic peptides exhibit 88.9-100% similarity to the TaqII sequence. All four enzymes were purified to homogeneity; their polypeptide sizes (114.5-122 kDa) make them the largest class-IIS restriction endonucleases known to date. The existence of a Thermus sp. sub-family of class-IIS restriction endonucleases of a common origin is herein proposed.