Production of a tributyltin-binding protein 2 knockout mutant strain of Japanese medaka, Oryzias latipes
Production of a tributyltin-binding protein 2 knockout mutant strain of Japanese medaka, Oryzias latipes
复制标题
日本青鳉三丁基锡结合蛋白2敲除突变株的制备
DOI：
10.1016/j.marpolbul.2020.111601
复制
发表时间：
2020
影响因子：
5.8
通讯作者：
Oshima Yuji
中科院分区：
文献类型：
--
作者：
Kato-Unoki Yoko;Takai Yuki;Nagano Yosuke;Matsunaga Satoshi;Enoki Shintaro;Takamura Takumi;Kim Sangwan;Kinoshita Masato;Kitano Takeshi;Shimasaki Yohei;Oshima Yuji
三丁基锡结合蛋白(TBT-Bps)是Lipocalin家族的成员，与鱼血中的TBT结合，被认为有助于TBT的解毒。最近的研究表明，许多鱼类物种都有TBT-BP基因，这些基因是由暴露在化学品或鱼类病原菌等应激条件下诱导的。然而，TBT-BPS的功能及其诱导和解毒作用的机制尚不清楚。在此，为了阐明Tbt-Bp2的功能，我们利用CRISPR/CAS9系统获得了日本青竹的Tbt-Bp2基因敲除株系(Tbt-Bp2−/−)。突变的TBT-bp2基因表达降低，基因序列测定和预测的蛋白质结构表明可能失去了功能。然而，这些鱼可以在正常条件下生长。在未来的实验中，将青竹的tbt-bp2−/−菌株暴露在各种压力下，有望有助于我们了解这种新的水生生物解毒系统。
Tributyltin-binding proteins (TBT-bps), members of the lipocalin family, bind TBT in fish blood and are presumed to contribute to detoxification of TBT. Recent studies have shown that many fish species have TBT-bp genes, and that these genes are induced by stresses such as exposure to chemicals or fish pathogenic bacteria. However, the function of TBT-bps, and the mechanisms of their induction and detoxification activity are still unclear. Here, towards elucidating the functions of TBT-bp2, we produced a TBT-bp2 knockout (TBT-bp2−/−) strain of Japanese medaka,Oryzias latipes, by using the CRISPR/Cas9 system. Gene expression of the mutated TBT-bp2 was reduced, and the cDNA sequencing and predicted protein structure suggested possible loss of function. However, the fish could be grown under normal conditions. Exposure of the TBT-bp2−/−strain of medaka to various stresses in future experiments is expected to contribute to our understanding of this novel detoxification system in aquatic organisms.