引入的构建体是通过CRISPR/CAS9系统诱导基因靶向敲除突变体的质粒载体。根据Smedley等人组装该质粒载体。,2021通过使用金门(GG,类型IIS限制酶)模块化克隆(MOCLO)组件。用于针对每个亚基因组中感兴趣基因的三个同源副本(a,b和d),在三个亚基因组的第一个外显子中选择了两个单个引导RNA(SGRNA)序列(sgrNA)序列(guide_2和gude_3)。如Smedley等人所述,使用WheatCrispr工具用于选择SGRNA。,2021。考虑了预测的目标和脱靶切割效率;预计SGRNA脱离目标得分的值为“ 0”,相当于根据WheatCrispr工具在其他遗传或基因间区域中的“无预测命中”。
一般性声明……………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………R4热转化…………………………………………………………………………………………………………………………………………………………………………………… R10责任………………………………………………………………………………………………………………………………………………………………………………………………………………………………………… R20的服务中断和削减服务………………………………………………………………………………………………………………………………………… R21帐单和付款………………………………………………………………………………………………………………………………………………………………………………………… R24隐私,访问和披露………………………………………………………………………………………………………………………………………………………… R27线扩展名………………………………………………………………………………………………………………………………………………………………………………………………………………………… R40注册,计量失败和其他计费错误的不准确性…………………………。 R52公司的检查……………………………………………………………………………………………………………………………………………………………………… R54客户存款…………………………………………………………………………………………………………………………………………………… R57服务停产………………………………………………………………………………………………………………………………………………………………………………………………………… R61停产通知…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………。 R70预算计费计划和分期付款………………………………………………………………………………………………………………………… R71天然气的转移……………………………………………………………………………………………………………………………………………………………………………………………………………… R75R4热转化……………………………………………………………………………………………………………………………………………………………………………………R10责任…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………R20的服务中断和削减服务………………………………………………………………………………………………………………………………………… R21帐单和付款………………………………………………………………………………………………………………………………………………………………………………………… R24隐私,访问和披露………………………………………………………………………………………………………………………………………………………… R27线扩展名………………………………………………………………………………………………………………………………………………………………………………………………………………………… R40注册,计量失败和其他计费错误的不准确性…………………………。 R52公司的检查……………………………………………………………………………………………………………………………………………………………………… R54客户存款…………………………………………………………………………………………………………………………………………………… R57服务停产………………………………………………………………………………………………………………………………………………………………………………………………………… R61停产通知…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………。 R70预算计费计划和分期付款………………………………………………………………………………………………………………………… R71天然气的转移……………………………………………………………………………………………………………………………………………………………………………………………………………… R75R20的服务中断和削减服务…………………………………………………………………………………………………………………………………………R21帐单和付款………………………………………………………………………………………………………………………………………………………………………………………… R24隐私,访问和披露………………………………………………………………………………………………………………………………………………………… R27线扩展名………………………………………………………………………………………………………………………………………………………………………………………………………………………… R40注册,计量失败和其他计费错误的不准确性…………………………。 R52公司的检查……………………………………………………………………………………………………………………………………………………………………… R54客户存款…………………………………………………………………………………………………………………………………………………… R57服务停产………………………………………………………………………………………………………………………………………………………………………………………………………… R61停产通知…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………。 R70预算计费计划和分期付款………………………………………………………………………………………………………………………… R71天然气的转移……………………………………………………………………………………………………………………………………………………………………………………………………………… R75R21帐单和付款…………………………………………………………………………………………………………………………………………………………………………………………R24隐私,访问和披露…………………………………………………………………………………………………………………………………………………………R27线扩展名………………………………………………………………………………………………………………………………………………………………………………………………………………………… R40注册,计量失败和其他计费错误的不准确性…………………………。 R52公司的检查……………………………………………………………………………………………………………………………………………………………………… R54客户存款…………………………………………………………………………………………………………………………………………………… R57服务停产………………………………………………………………………………………………………………………………………………………………………………………………………… R61停产通知…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………。 R70预算计费计划和分期付款………………………………………………………………………………………………………………………… R71天然气的转移……………………………………………………………………………………………………………………………………………………………………………………………………………… R75R27线扩展名…………………………………………………………………………………………………………………………………………………………………………………………………………………………R40注册,计量失败和其他计费错误的不准确性…………………………。R52公司的检查………………………………………………………………………………………………………………………………………………………………………R54客户存款……………………………………………………………………………………………………………………………………………………R57服务停产…………………………………………………………………………………………………………………………………………………………………………………………………………R61停产通知…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………。 R70预算计费计划和分期付款………………………………………………………………………………………………………………………… R71天然气的转移……………………………………………………………………………………………………………………………………………………………………………………………………………… R75R61停产通知…………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………………。R70预算计费计划和分期付款………………………………………………………………………………………………………………………… R71天然气的转移……………………………………………………………………………………………………………………………………………………………………………………………………………… R75R70预算计费计划和分期付款…………………………………………………………………………………………………………………………R71天然气的转移………………………………………………………………………………………………………………………………………………………………………………………………………………R75R75
