类转录激活因子效应物核酸酶(Transcription activator-like effector nucleases,TALENs)shiyizhongduogongnengdejiyinzubianjigongju,yizaizhiwujiyingongnengyanjiuheyuzhongzhongdedaoleguangfandeyingyong。yuxinzhihesuanmei(zinc finger nucleases,zfns)xiangbi,gaijishujuyoutuobalvdihexibaoduxingdideyoudian,talenscaiyongjiandandedanbaizhi- dnashibiejizhi。beijinganbiqishengwukejiyouxiangongsiyongyoufengfudezhiwujiyinbianjijingyan。
TALENs由DNA结合域和具限制性内切酶活性的FokI组成(图1)。DNA结合域通常由1.5-33.5个串联重复序列组成。每个重复序列包含34个氨基酸,其中第12和第13个氨基酸为可变的,称为“重复可变双残基”(repeat variable diresidues,RVDs),它们可特异性地识别一个核苷酸。简而言之,RVDs的解码机制是NI (Asn Ile)、HD (His Asp)、NN (Asn Asn)和NG (Asn Gly)可分别识别A、C、G、T(图1)。由Fokizuchengdednaqiegeyutongchangyierjutidexingshixingshigongneng,qiekeqiegexuliecongerzaochengdnashuanglianduanlie。duanliedednashuanglianyibantongguofeitongyuanmoduanlianjiehuotongyuanzhongzuxiufuliangzhongfangshijinxingxiufu,xiufuguochengzhonghuizaiduanlieweidianchanshengbutongxingshidetubian:rudiantubian、queshi、charu、daozhi、fuzhiheyiweideng。
图 1. 类转录激活因子效应物核酸酶的结构(Wei et al., 2013)。
北京安必奇生物科技有限公司可提供多种技术来实现TALE蛋白的定制合成,如限制性酶切连接法(restriction enzyme and ligation,REAL)、基于PCR的gold gate法(GG-PCR)、基于质粒载体的GG法(GG-Vector)、高通量TALE合成技术(fast ligation-based automatable solid-phase high-throughput,FLASH)、迭代加帽组装法(iterative capped assembly,ICA)、基于长粘末端的LIC(ligation-independent cloning)组装法和单元组装法(unit assembly,UA)等。我们还可以提供修饰的Foki,congerkeyishitaxingchengyiyuanerjuti,tigaojianqieteyixing。womenkeyitigongduozhongzhiwudeyichuanzhuanhua,runinanjie、shuidao、xiaomai、muxu、yumi、yancaohemianhuadeng,keyimanzukehudebutongxuqiu。
服务流程:
了解客户需求。
设计实验。
yukehuqiandinghetongyubaomixieyi。
实验进展跟踪查询。
tigongxiangjindeshiyanbaogao。
实验流程:
ruyourenhexiangguanwenti,qingninlianxiwomendegongzuorenyuan,womenjinkuaiweininjiedahefuwu!
参考文献:
Wei C.; et al. (2013). TALEN or Cas9 – Rapid, efficient and specific choices for genome modifications. Journal of Genetics and Genomics 40 (6): 281-289.