芒果SEPALAATA基因的克隆与表达分析

以芒果品种‘吕宋’(Mangifera indica L.Carabao)为试材,利用同源克隆和RACE技术从花序中获得了1个芒果SEPALAATA(SEP)基因cDNA全长,命名为MSEP1(GenBank中登录号为KP702299)。MSEP1基因的cDNA全长为921bp,包含一个长度为726bp开放阅读框,编码241个氨基酸,蛋白质相对分子质量为27.7kD,理论等电点为5.79。序列比对和系统进化树分析表明,MSEP1具有保守的MADS-box及半保守的K区,属于MADS-box家族的SEP亚家族。器官特异性表达分析表明,MSEP1基因在芒果根、茎中表达量较低,在叶片、花芽中表达量较高,而在花序中表达量最高。研究推测,MSEP1基因可能在芒果生殖生长中发挥重要作用。     

Induction of small-segment-translocation between wheat and rye chromosomes

A new approach to produce wheat-rye translocation, based on the genetic instability caused by monosomic addition of rye chromosome in wheat, is described. 1 283 plants from the selfed progenies of monosomic addition lines with single chromosome of inbred rye line R12 and complete chromosome complement of wheat cultivar Mianyang 11 were cytologically analyzed on a plant-by-plant basis by the improved C-banding technique. 63 of the plants, with 2n = 42, were found containing wheat-rye translocation or substitution, with a frequency of 4. 91% . Compared with the wheat parent, other 32 plants with 2n = 42 exhibited obvious phenotypic variation, but their com-ponent of rye chromosome could not be detected using the C-banding technique. In situ hybridization with a biotin-la-beled DNA probe was used to detect rye chromatin and to determine the insertion sites of rye segments in the wheat chromosomes. In 20 out of the 32 variant wheat plants, small segments of rye chromosomes were found being inserted into dif     

斑痣盘菌科一新种——松生小鞋孢盘菌

本文报道在云南省昆明市华山松(Pinus armandi Franch.)上发现的斑痣盘菌科一新种,即松生小鞋孢盘菌(Soleella pinicola Y.R.Lin et W.Ren)。对该种作了拉丁文、汉文描述和图解。主模式标本保藏于安徽农学院林学系森林保护教研室。     

热带假丝酵母1230POX4、POX5基因及其编码蛋白的研究

根据GenBank中热带假丝酵母菌株pk233(ATCC 20336)POX4和POX5基因序列,设计了两对引物。通过PCR扩增,用一种简易的方法克隆了1230菌株中这两个基因。对POX4、POX5的测序表明,1230菌株的POX5基因与pk233菌株的POX5基因存在菌株间的差异。这种差异对蛋白功能的影响有待进一步研究。对POX4和POX5编码的蛋白PXP4、PXP5进行Pfam分析和二级结构预测后,推测PXP4和PXP5的N端可能是FAD结合部位。     

基因前定点敲入FLAG标签表达的FLAG-SND1蛋白参与胞内应激颗粒的形成

CRISPR/ Cas9 (clustered regular interspaced short palindromic repeats-Cas9 nuclease) gene editing technology is an emerging technology for gene-specific knock-out and knock-in in recent years. In this experiment, the CRISPR/ Cas9 gene editing system was used to insert 3×FLAG tags into the front of SND1 gene in HeLa cells, so that the endogenous expression of SND1 protein in cells was equipped with 3×FLAG tags, and the localization of SND1 with stress granules and processing bodies was observed. A sgRNA near the start codon ATG of the SND1 gene was designed, and a recombinant eukaryotic expression plasmid was constructed using px459 as an expression vector. A sequence containing 3×FLAG and a 150 bp homologous arm upstream and downstream of the position to be inserted was designed, and the recombinant plasmid was synthesized by the company. Two plasmids were co-transfected into HeLa cells, and positive cells were screened by puromycin. Western blotting indicated that the cells expressed the 3×FLAG-SND1 fusion protein. Genomic DNA was extracted and sequenced. Cell cycle and apoptosis were detected by flow cytometry after sequencing and the stable strain was obtained without error. It was found that there was no significant difference compared with WT cells. At the same time, the treatment with 0. 5 mmol / L sodium arsenite resulted in oxidative stress in cells, increased phosphorylation of eIF2α protein, and the presence of stress particles in the cytoplasm. There was co-localization between SND1 and stress particle marker protein TIAR, but no co-localization with processed body protein DCP1α. © 2019 The authors.     

悬浮培养发状念珠蓝细菌对盐碱胁迫的生理应答研究北大核心CSCD

天然发状念珠蓝细菌是荒漠化土壤中的"先锋生物",具有极强的耐旱、耐碱、耐高温等能力。本研究旨在揭示悬浮培养发状念珠蓝细菌的抗盐碱能力,为发状念珠蓝细菌的人工培养提供基础。25℃,80μmol/(m2·s)光照下,BG-11培养液培养发状念珠蓝细菌,利用不同浓度(0、60、120、180、240 mmol/L)混合的Na2SO4和Na2CO3胁迫发状念珠蓝细菌细胞,处理时间分别为24 h和72 h,测定其质膜透性、超氧化物歧化酶活性、丙二醛、脯氨酸、可溶性蛋白以及可溶性多糖含量,分析发状念珠蓝细菌细胞对盐碱胁迫的响应。盐碱胁迫下,随着胁迫程度的加重,发状念珠蓝细菌细胞的质膜相对透性增加,丙二醛含量上升;超氧化物歧化酶活性和脯氨酸含量先上升后下降;可溶性蛋白含量逐渐下降;可溶性多糖含量不断升高。悬浮培养的发状念珠蓝细菌细胞对盐碱胁迫产生结构和生理的应激响应,增强了发状念珠蓝细菌抗盐碱能力。     

Dega骨盆截骨术后最佳中心边缘角的三维有限元分析

目的:应用三维有限元技术评估发育性髋关节脱位(developmental dysplasia of the hip,DDH)患儿在Dega骨盆截骨术后不同中心边缘角(center-edge angle,CEA)状态下髋臼的应力分布,为术前的手术规划提供有参考价值的生物力学结果。方法:使用已建立的DDH患者髋关节三维有限元模型,以术后CEA27°为中间值,每3°为一个变量,在Mimics誖10.0软件的模拟手术模块分别构建7组髋臼截骨术后的模型。在单腿站立和双腿站立状态下测量不同CEA状态下髋臼的应力分布。结果:单腿站立情况下,CEA为24°、27°、30°和33°的术后模型患侧髋臼的峰值应力接近正常侧。双腿站立情况下,CEA为24°时双侧髋臼的峰值应力最为接近。结论:对于该患者而言,在7组术后CEA中,24°时患侧髋臼的峰值应力与健侧最为接近,可以认为是最佳的术后CEA。有限元技术能够为Dega手术的术前规划提供个性化的指导方案。     

乏氧和辐射诱导的miR-1290 促进宫颈癌细胞EMT发生

目的:研究mi R-1290在宫颈癌Hela细胞上皮间质转化中的作用。方法:模拟肿瘤放射治疗的分割疗法,单次2 Gyγ射线连续照射宫颈癌Hela、Siha细胞诱导辐射抗性细胞株;采用micro RNA芯片技术比较辐射抗性细胞与亲本细胞中mi RNA的表达谱差异;经不同条件乏氧和辐射处理宫颈癌Hela细胞后检测mi R-1290的表达水平;借助mi R-1290及mi R-1290-inhibition慢病毒表达载体,调变mi R-1290在Hela细胞的表达;调变mi R-1290后,划痕实验及Transwell侵袭实验比较Hela细胞的侵袭和转移能力;蛋白质印迹法检测细胞中E钙黏素(E-cadherin)和N钙黏素(N-cadherin)的表达。结果:在宫颈癌辐射抗性细胞中mi R-1290表达水平显著升高(P0.05);乏氧和辐射可诱导mi R-1290在宫颈癌Hela细胞中表达(P0.05);上调表达mi R-1290,Hela细胞出现了明显的间质细胞的形态改变,细胞的侵袭和转移能力明显增强(P0.05)。在Hela细胞中上调表达mi R-1290,降低了细胞中E-cadherin的表达,同时升高了N-cadherin的表达(P0.05)。结论:乏氧和辐射可诱导mi R-1290在宫颈癌Hela细胞中表达,mi R-1290通过调控E-cadherin和N-cadherin的表达促进宫颈癌Hela细胞发生EMT。     

转双基因抗虫棉对棉铃虫的抗性

将苏云金杆菌 (Ｂａｃｉｌｌｕｓｔｈｕｒｉｎｇｉｅｎｓｉｓ,Ｂｔ)杀虫晶体蛋白 (ｉｎｓｅｃｔｉｃｉｄａｌｃｒｙｓｔａｌｐｒｏｔｅｉｎ ,ＩＣＰ)基因转入棉花植株中获得的抗虫棉花 (下略为Ｂｔ棉 ) ,为棉花害虫综合防治开辟了新的途径。Ｂｔ棉除具有丰产性、纤维品质好和对靶标害虫的良好控制作用等特性外 ,更重要的应用价值在于其高杀虫效果与维护生态环境的协调发展的良性作用上[1] 。但研究结果已表明 ,害虫对ＢｔＩＣＰ的抗性适应 ,将严重威胁Ｂｔ棉的使用寿命[2 ] 。已有的报道证明 ,将两种不同作用机制的杀虫基因转入植物并同时表…