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马铃薯StZIP8–like基因的克隆及镉转运功能分析
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作者 杨丽红 席银凯 +1 位作者 王丹 胡新喜 《湖南农业大学学报(自然科学版)》 北大核心 2025年第3期16-23,共8页
为了探讨StZIP8–like基因的表达模式和功能,从马铃薯中克隆得到StZIP8–like基因的全长序列,通过生物信息学方法对该基因的结构、理化性质及亲缘关系进行预测;利用RT–qPCR技术,分析其在镉胁迫下的表达模式;构建融合表达载体,分析StZIP... 为了探讨StZIP8–like基因的表达模式和功能,从马铃薯中克隆得到StZIP8–like基因的全长序列,通过生物信息学方法对该基因的结构、理化性质及亲缘关系进行预测;利用RT–qPCR技术,分析其在镉胁迫下的表达模式;构建融合表达载体,分析StZIP8–like蛋白在烟草叶片中的亚细胞定位;通过酵母异源表达试验,分析St ZIP8–like的镉转运功能。结构分析结果表明,StZIP8–like序列全长1 044 bp,编码347个氨基酸,含有1个ZIP保守结构域及7个跨膜结构域;系统进化分析结果表明,StZIP8–like蛋白与番茄及烟草中的ZIP8蛋白的亲缘关系较近。亚细胞定位结果显示,St ZIP8–like蛋白定位于细胞质膜;RT–q PCR结果表明,St ZIP8–like在马铃薯幼苗的根中表达量较高,并且Cd胁迫诱导下St ZIP8–like基因在马铃薯根、茎和叶中的表达量均上调;酵母试验结果显示,St ZIP8–like过表达的酵母对Cd更加敏感,具有Cd吸收转运活性。以上结果说明St ZIP8–like受到Cd胁迫的诱导表达,可能参与马铃薯吸收Cd的过程。 展开更多
关键词 马铃薯 stzip8–like 基因表达 酵母
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抗虫基因Cry8-like在大豆中的遗传转化及抗虫性初步鉴定 被引量:2
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作者 林楠 吴楠 +7 位作者 冯咏琪 于淼 戎洁 敖振超 闫鸽 姚丹 曲静 王丕武 《中国油料作物学报》 CAS CSCD 北大核心 2017年第2期221-227,共7页
利用无缝克隆技术把抗虫基因Cry8-like重组到pCAMBIA3301载体上,成功构建了含有抗草铵膦筛选标记的植物表达载体pCAMBIA3301-RSP-Cry8-like-nos。将该载体通过农杆菌介导法转入两个受体大豆品种吉农28和吉农42中,经PCR检测,吉农28和吉... 利用无缝克隆技术把抗虫基因Cry8-like重组到pCAMBIA3301载体上,成功构建了含有抗草铵膦筛选标记的植物表达载体pCAMBIA3301-RSP-Cry8-like-nos。将该载体通过农杆菌介导法转入两个受体大豆品种吉农28和吉农42中,经PCR检测,吉农28和吉农42分别获得了17株和13株T_1阳性植株,36株和25株T_2阳性植株。T_2转基因植株Southern杂交结果显示,目的基因以单拷贝的形式整合到大豆基因组中。荧光定量PCR检测结果表明,Cry8-like在转基因植株幼根得到了表达,而非转化受体对照未检测到荧光信号。室内抗暗黑鳃金龟幼虫鉴定结果表明,转基因植株提高了抗暗黑鳃金龟幼虫的能力。 展开更多
关键词 大豆 Cry8-like基因 根系特异表达载体 抗虫性
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家蚕BmCaspase-8-Like(BmCasp8L)的免疫负调控功能 被引量:1
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作者 胡杰 王鑫怡 王菲 《中国农业科学》 CAS CSCD 北大核心 2018年第21期4188-4196,共9页
【目的】在个体和细胞水平上探讨家蚕(Bombyx mori)BmCaspase-8-Like(BmCasp8L)的免疫负调控功能,为研究昆虫免疫负调控机制提供参考。【方法】通过RT-PCR技术克隆BmCasp8L并进行结构域预测和进化分析;利用荧光定量PCR检测BmCasp8L在家... 【目的】在个体和细胞水平上探讨家蚕(Bombyx mori)BmCaspase-8-Like(BmCasp8L)的免疫负调控功能,为研究昆虫免疫负调控机制提供参考。【方法】通过RT-PCR技术克隆BmCasp8L并进行结构域预测和进化分析;利用荧光定量PCR检测BmCasp8L在家蚕各龄期、5龄第3天及预蛹期各组织中的时空表达特征和家蚕感染细菌后的免疫诱导表达特征;合成用于RNAi的dsRNA,通过注射dsRNA在个体中降低BmCasp8L的表达水平,并检测对抗菌肽基因表达水平的影响;构建BmCasp8L的细胞表达载体,通过质粒或dsRNA的转染在BmE细胞中过表达BmCasp8L或降低BmCasp8L的表达水平,并利用Western blot或定量PCR予以确认,同时检测抗菌肽基因表达水平的变化及转录因子BmRelish的切割。【结果】BmCasp8L与鳞翅目Caspase-6、哺乳动物Caspase-8同源,其序列与BmDredd、DmDredd N端分别具有61%、42%的相似性,但缺少C端的Caspase结构域。时空表达特征分析表明,BmCasp8L在眠蚕期表达量高于起蚕期,在预蛹期、蛹第7天、蛾第1天表达量明显升高;在5龄第3天幼虫体内,BmCasp8L在血细胞中的表达量明显高于其他组织,而在预蛹期,BmCasp8L主要在丝腺中表达。免疫诱导表达谱显示,5龄第3天家蚕在注射感染黑胸败血芽孢杆菌或粘质沙雷氏菌1 h内,BmCasp8L的表达水平上升,此后逐渐恢复正常。对5龄第2天家蚕注射dsCasp8L或dsEGFP,24 h后通过荧光定量PCR检测到BmCasp8L的表达量在注射dsCasp8L的家蚕中显著下调,而抗菌肽BmCecropinA1的表达水平显著上调。在BmE细胞中过表达BmCasp8L,抗菌肽BmCecropinA1的表达水平与对照相比显著降低,且转录因子BmRelish的切割受到抑制。在细胞中转染dsRNA后,BmCasp8L的表达水平被有效降低,抗菌肽BmCecropinA1的表达水平显著上调。【结论】进化分析、表达特征以及细胞和个体上的功能研究表明BmCasp8L是一个具有免疫负调控作用的分子,通过抑制BmRelish的切割,抑制抗菌肽的表达,从而负调控Imd信号通路,降低BmCasp8L的表达水平则导致抗菌肽表达水平升高,有助于家蚕抵御微生物病原的侵染。 展开更多
关键词 家蚕 免疫负调控 BmCaspase-8-like(BmCasp8L) 表达特征 抗菌肽 BmRelish
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Design of 8-like Non-planar Cavity 被引量:1
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作者 YANG Jian-qiang 《Semiconductor Photonics and Technology》 CAS 2008年第4期229-233,共5页
For eliminating the negative effects of lock-in region and optical elements in cavity,the non-planar structure is strongly needed in laser gyro. In this paper,the theoretical formula of rotatory effect in non-planar c... For eliminating the negative effects of lock-in region and optical elements in cavity,the non-planar structure is strongly needed in laser gyro. In this paper,the theoretical formula of rotatory effect in non-planar cavity is presented deducing from basic light propagating law. Based on the 8-like plane cavity,a novel non-planar structure is designed and numerically calculated. The results show that for obtaining the required rotatory angle,it just needs to adjust the structural parameters of the original 8-like plane cavity. The four-frequency-differential laser gyro which adopts the non-planar 8-like structure has the same advantages as those with the planar 8-like structure,such as the gain region locates at one leg,the position of Faraday rotator has better spatial symmetry and the coating films of the output mirror is easily designed. 展开更多
关键词 LASERS non-planar cavity 8-like plane cavity rotatory effect
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Pax-8基因敲除小鼠心脏中Bcl2l14基因表达上调 被引量:3
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作者 高瞻 来丹丹 +4 位作者 黄晓燕 褚茂平 施翔翔 张怀勤 杨德业 《中国病理生理杂志》 CAS CSCD 北大核心 2010年第4期640-644,共5页
目的:寻找先天性心脏病相关基因-转录因子Pax-8的下游基因。方法:分别提取Pax-8基因敲除小鼠纯合子(Pax-8 KO-/-)和杂合子(Pax-8 KO+/-)的心脏总RNA,利用含31802个小鼠基因的基因芯片检测两组小鼠基因表达水平,找出差异表达的基因,并经... 目的:寻找先天性心脏病相关基因-转录因子Pax-8的下游基因。方法:分别提取Pax-8基因敲除小鼠纯合子(Pax-8 KO-/-)和杂合子(Pax-8 KO+/-)的心脏总RNA,利用含31802个小鼠基因的基因芯片检测两组小鼠基因表达水平,找出差异表达的基因,并经半定量RT-PCR和荧光实时定量PCR技术初步筛选出转录因子Pax-8的下游基因。结果:基因芯片检测发现,Pax-8 KO-/-组与Pax-8 KO+/-相比有25个基因表达下调,另有17个基因表达上调,差异基因涉及细胞周期及信号转导的调节因子,直接参与代谢的酶,以及核转录因子等。用半定量RT-PCR验证发现:Bcl2-like 14(Bcl2l14)基因在Pax-8 KO-/-组上调。定量RT-PCR亦证实在Pax-8KO-/-组Bcl2l14基因的表达水平较Pax-8 KO+/-组及Pax-8 KO+/+(野生型)组分别上调2.07倍和2.23倍(P<0.01)。结论:Bcl2l14基因为转录因子Pax-8的下游基因,可能在先天性心脏病室间隔缺损的发病机制中发挥重要作用。 展开更多
关键词 Paired BOX GENE 8 Bcl2-like 14 室间隔缺损 细胞凋亡
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Dyslipidaemia of diabetes and the intestine 被引量:2
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作者 Gerald H Tomkin Daphne Owens 《World Journal of Diabetes》 SCIE CAS 2015年第7期970-977,共8页
Atherosclerosis is the major complication of diabetes and has become a major issue in the provision of medical care.In particular the economic burden is growing at an alarming rate in parallel with the increasing worl... Atherosclerosis is the major complication of diabetes and has become a major issue in the provision of medical care.In particular the economic burden is growing at an alarming rate in parallel with the increasing worldwide prevalence of diabetes.The major disturbance of lipid metabolism in diabetes relates to the effect of insulin on fat metabolism.Raised triglycerides being the hallmark of uncontrolled diabetes,i.e.,in the presence of hyperglycaemia.The explosion of type 2 diabetes has generated increasing interest on the aetiology ofatherosclerosis in diabetic patients.The importance of the atherogenic properties of triglyceride rich lipoproteins has only recently been recognised by the majority of diabetologists and cardiologists even though experimental evidence has been strong for many years.In the post-prandial phase 50% of triglyceride rich lipoproteins come from chylomicrons produced in the intestine.Recent evidence has secured the chylomicron as a major player in the atherogenic process.In diabetes chylomicron production is increased through disturbance in cholesterol absorption,in particular Neimann Pick C1-like1 activity is increased as is intestinal synthesis of cholesterol through 3-hydroxy-3-methyl glutaryl co enzyme A reductase.ATP binding cassette proteins G5 and G8 which regulate cholesterol in the intestine is reduced leading to chylomicronaemia.The chylomicron particle itself is atherogenic but the increase in the triglyceride-rich lipoproteins lead to an atherogenic low density lipoprotein and low high density lipoprotein.The various steps in the absorption process and the disturbance in chylomicron synthesis are discussed. 展开更多
关键词 TRIGLYCERIDE Cholesterol CHYLOMICRONS Microsomal TRIGLYCERIDE transfer protein Niemann PickC1-like1 LIPOPROTEINS DIABETES ATP binding cassetteproteins G5/G8
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BmCaspase-8-like regulates autophagy by suppressing BmDREDD-mediated cleavage of BmATG6
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作者 Chang Sun Dongmei Wei +2 位作者 Yumeng Pan Xiaoyi Xiao Fei Wang 《Insect Science》 SCIE CAS CSCD 2023年第2期365-374,共10页
Autophagy plays an important role in tissue remodeling during insect development.The interplay between autophagy-related(ATG)proteins and caspases regulates the autophagic activity of ATGs,thereby modulating the proce... Autophagy plays an important role in tissue remodeling during insect development.The interplay between autophagy-related(ATG)proteins and caspases regulates the autophagic activity of ATGs,thereby modulating the process of autophagy.Our previous study characterized BmCaspase-8-like(BmCasp8L)as a caspase suppressor that inhibits apoptosis and immune signaling by suppressing the activation of death-related ced-3/Nedd2-like caspase(DREDD),a caspase-8 homolog in silkworm.In this study,we explored the regulatory role of BmCasp8L in autophagy.We found that the expression of Bmcasp8l increased from the late spinning stage to the pupa stage in the posterior silk gland(PSG),correlating with the expression patterns of Bmatg8 and Bmatg6.RNA interference-mediated downregulation of BmCasp8L expression significantly decreased starvation-induced autophagic influx as determined by the levels of BmATG8–phosphatidylethanolamine and the percentage of cells displaying punctate enhanced green fluorescent protein-BmATG8.Conversely,the overexpression of BmCasp8L significantly increased autophagic influx.We also found that BmCasp8L underwent autophagic degradation induced by starvation and that it was colocalized with BmATG8.Lastly,we demonstrated that BmDREDD attenuated autophagy and BmCasp8L suppressed BmDREDD-mediated cleavage of BmATG6.Taken together,our results demonstrated that BmCasp8L is a novel proautophagic molecule which suppresses BmDREDD-mediated cleavage of BmATG6 and is a target for autophagy. 展开更多
关键词 ATG6 ATG8 AUTOPHAGY Bombyx mori Caspase-8-like DREDD
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