Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321&#...Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.展开更多
目的绘制肠型胃癌的超级增强子重塑图谱,揭示超级增强子的肿瘤生物学功能及可能激活的下游靶基因。方法收集陆军特色医学中心消化内科2022年1-12月收治行内镜检查、治疗的患者的31例正常胃黏膜组织、23例肠型胃癌组织及9例肠型胃癌类器...目的绘制肠型胃癌的超级增强子重塑图谱,揭示超级增强子的肿瘤生物学功能及可能激活的下游靶基因。方法收集陆军特色医学中心消化内科2022年1-12月收治行内镜检查、治疗的患者的31例正常胃黏膜组织、23例肠型胃癌组织及9例肠型胃癌类器官进行靶向组蛋白H3K27ac修饰的染色质靶向捕获(cleavage under targets and tagmentation,CUT&Tag)测序,基于生物信息学工具描绘肠型胃癌的超级增强子重塑特征并鉴定关键靶基因;利用CRISPRi技术干预超级增强子,Western blot检测靶基因的表达,并采用CCK-8法和Transwell迁移、侵袭实验检测对照组细胞及干预组细胞的增殖、迁移、侵袭能力。结果肠型胃癌组织与正常胃黏膜组织超级增强子信号差异明显(P<0.05),癌组织中的活性超级增强子可能参与免疫系统的负向调节、细胞黏附等生物学过程;在肿瘤细胞中表达上调的细胞迁移诱导蛋白(cell migration-inducing protein,CEMIP)受到超级增强子的调控,干预超级增强子后肿瘤细胞CEMIP蛋白表达量下降(P<0.05),细胞增殖率、相对迁移面积和相对侵袭面积也随之下降(P<0.05)。结论肠型胃癌发生了超级增强子重塑,其下游靶基因CEMIP可促进癌细胞生长、迁移和侵袭。展开更多
【目的】基于猪极端饲料报酬表型,利用靶向切割标签技术(cleavage under targets and tagmentation,CUT&Tag)构建H3K27ac差异图谱,鉴定关联的启动子、增强子和关联候选基因,为猪饲料报酬解析提供基础数据。【方法】从209头杜洛克猪...【目的】基于猪极端饲料报酬表型,利用靶向切割标签技术(cleavage under targets and tagmentation,CUT&Tag)构建H3K27ac差异图谱,鉴定关联的启动子、增强子和关联候选基因,为猪饲料报酬解析提供基础数据。【方法】从209头杜洛克猪中筛选饲料报酬表型差异最大的4头个体分别组成低剩余采食量组(L组,n=2)和高剩余采食量组(H组,n=2)。利用CUT&Tag技术,以H3K27ac作为活性启动子和增强子的表观遗传标记对4头个体的肝脏组织进行全基因组扫描,鉴定关联启动子、增强子和候选基因,并进一步采用real-time quantitative PCR(RT-qPCR)对重要基因进行验证。【结果】共鉴定出47271个H3K27ac峰,包括15739个潜在启动子和31532个推定增强子,大部分的H3K27ac峰位于近端启动子区域。根据H3K27ac全基因的分布情况和作用特点,发现3087个差异峰(对应2188个基因),其中867个H3K27ac峰(对应664个基因)富集在H组,2220个H3K27ac峰(对应1575个基因)富集在L组。GO和KEGG结果发现H3K27ac增强子可能是通过影响胆汁酸代谢、淋巴细胞代谢和NF-κB通路来调节饲料报酬。RT-qPCR结果显示,与H组相比,DIAPH3、DPYD、FTO、TCF7L2、ABCC2、ABCC11、RXRA和ABCG8在L组中显著上调(P<0.05),而PAX5、GRHPR、NFATC1、CARD11、BLNK和LYN在L组中显著下调(P<0.05)。【结论】本研究成功生成了具有极端RFI差异的杜洛克猪肝脏组织中的H3K27ac差异图谱以及相关数据。基于该图谱鉴定和验证了关联候选基因,为进一步解析启动子和增强子调控饲料报酬的分子机制提供了基础数据。展开更多
文摘Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.
文摘目的绘制肠型胃癌的超级增强子重塑图谱,揭示超级增强子的肿瘤生物学功能及可能激活的下游靶基因。方法收集陆军特色医学中心消化内科2022年1-12月收治行内镜检查、治疗的患者的31例正常胃黏膜组织、23例肠型胃癌组织及9例肠型胃癌类器官进行靶向组蛋白H3K27ac修饰的染色质靶向捕获(cleavage under targets and tagmentation,CUT&Tag)测序,基于生物信息学工具描绘肠型胃癌的超级增强子重塑特征并鉴定关键靶基因;利用CRISPRi技术干预超级增强子,Western blot检测靶基因的表达,并采用CCK-8法和Transwell迁移、侵袭实验检测对照组细胞及干预组细胞的增殖、迁移、侵袭能力。结果肠型胃癌组织与正常胃黏膜组织超级增强子信号差异明显(P<0.05),癌组织中的活性超级增强子可能参与免疫系统的负向调节、细胞黏附等生物学过程;在肿瘤细胞中表达上调的细胞迁移诱导蛋白(cell migration-inducing protein,CEMIP)受到超级增强子的调控,干预超级增强子后肿瘤细胞CEMIP蛋白表达量下降(P<0.05),细胞增殖率、相对迁移面积和相对侵袭面积也随之下降(P<0.05)。结论肠型胃癌发生了超级增强子重塑,其下游靶基因CEMIP可促进癌细胞生长、迁移和侵袭。
文摘【目的】基于猪极端饲料报酬表型,利用靶向切割标签技术(cleavage under targets and tagmentation,CUT&Tag)构建H3K27ac差异图谱,鉴定关联的启动子、增强子和关联候选基因,为猪饲料报酬解析提供基础数据。【方法】从209头杜洛克猪中筛选饲料报酬表型差异最大的4头个体分别组成低剩余采食量组(L组,n=2)和高剩余采食量组(H组,n=2)。利用CUT&Tag技术,以H3K27ac作为活性启动子和增强子的表观遗传标记对4头个体的肝脏组织进行全基因组扫描,鉴定关联启动子、增强子和候选基因,并进一步采用real-time quantitative PCR(RT-qPCR)对重要基因进行验证。【结果】共鉴定出47271个H3K27ac峰,包括15739个潜在启动子和31532个推定增强子,大部分的H3K27ac峰位于近端启动子区域。根据H3K27ac全基因的分布情况和作用特点,发现3087个差异峰(对应2188个基因),其中867个H3K27ac峰(对应664个基因)富集在H组,2220个H3K27ac峰(对应1575个基因)富集在L组。GO和KEGG结果发现H3K27ac增强子可能是通过影响胆汁酸代谢、淋巴细胞代谢和NF-κB通路来调节饲料报酬。RT-qPCR结果显示,与H组相比,DIAPH3、DPYD、FTO、TCF7L2、ABCC2、ABCC11、RXRA和ABCG8在L组中显著上调(P<0.05),而PAX5、GRHPR、NFATC1、CARD11、BLNK和LYN在L组中显著下调(P<0.05)。【结论】本研究成功生成了具有极端RFI差异的杜洛克猪肝脏组织中的H3K27ac差异图谱以及相关数据。基于该图谱鉴定和验证了关联候选基因,为进一步解析启动子和增强子调控饲料报酬的分子机制提供了基础数据。
