Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity     

Anguo Li  Ruihao Huang  Chaogang Wang  Qunju Hu  Hui Li  Xiao Li 《Marine drugs》2021,19(5)

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.  相似文献   

3种染色体组加倍方法诱导黄河鲇雌核发育的比较          下载免费PDF全文

彭新亮  李学军  程果  乔志刚  石灵 《水生态学杂志》2008,29(2):44-47

用经紫外线照射后遗传失活的黄河鲇精子、刺激性成熟的黄河鲇卵子, 进行雌核发育, 采用冷休克、热休克和秋水仙素浸泡等3种染色体组加倍方法诱导黄河鲇卵进行二倍体雌核发育。实验结果表明: 在人工诱导黄河鲇二倍体雌核发育过程中, 以冷休克处理组的效果最好, 胚胎孵化率达到8. 1%; 其次为秋水仙素处理组, 胚胎孵化率达到6. 3%; 热休克处理组诱导效果较差, 胚胎孵化率仅为4. 0%。  相似文献   

拟无枝酸菌宿主构建及次级代谢基因簇异源表达     

胡欣瑞  贺卫军  吕金  王业民  陶美凤 《华中农业大学学报》2022,41(3):164-172

为将生长快、发酵时间短、遗传操作便捷的稀有放线菌拟无枝酸菌TNS106开发成异源表达宿主,通过同源重组将内源的瑞斯托霉素生物合成关键基因rpsA替换为ΦC31和ΦBT1噬菌体细菌附着位点attB,清除代谢背景并引入整合位点,得到菌株HXR1;将含有来自天蓝色链霉菌的放线紫红素基因簇或来自刺糖多孢菌的多杀菌素基因簇的质粒转到HXR1进行异源表达,检测发酵产物。结果显示,HXR1成功表达放线紫红素和多杀菌素;与红色糖多孢菌宿主LJ161相比,放线紫红素的产生提前1 d,产量提高1.3倍。拟无枝酸菌异源表达宿主HXR1可为从链霉菌和稀有放线菌中发现新的次级代谢产物提供有用的平台。  相似文献   

栉孔扇贝人工雌核发育的细胞学观察   总被引：5，自引：1，他引：5  

潘英 《水产学报》2004,28(6):616-622

运用荧光显微镜观察了栉孔扇贝正常卵子与雌核发育卵子在受精和成熟分裂过程中的核相变化。结果表明,尽管紫外线照射没有影响受精卵的成熟分裂以及雌性、雄性原核的形成,但使雌核卵的发育速度出现明显的滞缓。在第1次卵裂中期,雌核发育卵子中的雄性原核没有像雌性原核那样形成染色体,而是浓缩为一染色质小体(DCB),游离在细胞质中,不参与核分裂。胞质分裂结束时,DCB位于2个卵裂球之一的细胞质内或在赤道板处被分割成两部分。实验结果首次提供了栉孔扇贝雌核发育的细胞学证据。  相似文献   

人工诱导海水鱼类雌核发育的研究进展     

胡则辉  徐君卓 《海洋渔业》2007,29(1):78-83

近三十年来,国内外海水鱼类的增养殖业发展迅速,尤其以提高增养殖鱼类产量和质量为主要目的的染色体组工程已成为海水鱼类育种的热点.作为一种实用的染色体组操作技术,雌核发育是在雌核控制下的发育,精子只作为"激活源",入卵后不形成雄核,所以后代的遗传特质仅仅来源于母性亲本.  相似文献   

诱导栉孔扇贝雌核发育时精子入卵的扫描电镜观察   总被引：3，自引：0，他引：3       下载免费PDF全文

赵峰  杨爱国  刘志鸿  周丽青  王清印 《中国水产科学》2004,11(4):281-285

取栉孔扇贝 (Chlamysfarreri)精子在 80 0 μW/(cm2 ·s)的紫外线下辐射 ,然后与正常卵子受精 ,每隔 0 .5min取“受精卵”固定。在扫描电镜下连续观察精子的入卵过程 ,然后与正常精子的入卵过程进行比较。结果表明 ,紫外辐射后的精子可以划分为 3种类型 :I类精子 ,基本无明显变化 ,可以激发卵黄膜举起形成受精膜 ,同时诱发卵子发生皮层反应 ,在受精锥的作用下正常入卵 ,但入卵的时间较正常精子晚 ;II类精子 ,顶体膜破裂 ,顶体丝伸出 ;III类精子 ,顶体丝伸出 ,鞭毛脱落。由于II、III类精子在紫外辐射条件下诱发了顶体反应 ,融解卵膜的酶提前释放 ,因而不能入卵。精子鞭毛的脱落 ,使精子丧失了运动能力 ,是导致卵表面附着的精子数量较少的原因之一。无论是正常精子还是辐射后的精子 ,入卵后在卵表面都留下一个类似受精孔的通道。实验组中未发现有多精入卵现象  相似文献   

Production of Avaroferrin and Putrebactin by Heterologous Expression of a Deep-Sea Metagenomic DNA     

Masaki J. Fujita  Ryuichi Sakai 《Marine drugs》2014,12(9):4799-4809

The siderophore avaroferrin (1), an inhibitor of Vibrio swarming that was recently identified in Shewanella algae B516, was produced by heterologous expression of the biosynthetic gene cluster cloned from a deep-sea sediment metagenomic DNA, together with two analogues, bisucaberin (2) and putrebactin (3). Avaroferrin (1) is a macrocyclic heterodimer of N-hydroxy-N-succinyl cadaverine (4) and N-hydroxy-N-succinyl-putrescine (5), whereas analogues 2 and 3 are homodimers of 4 and 5, respectively. Heterologous expression of two other related genes from culturable marine bacteria resulted in production of compounds 1–3, but in quite different proportions compared with production through expression of the metagenomic DNA.  相似文献   

多尺度分解双寻优策略SPCNN的果园苹果异源图像融合模型     

刘立群  顾任远  周煜博  火久元 《农业工程学报》2022,38(17):158-167

针对单一的自然场景图像信息不能满足准确识别果实和精准定位目标的要求,提出一种多尺度分解双寻优策略简化脉冲耦合神经网络（Simplified Pulse Coupled Neural Network, SPCNN）的飞行时间（Time of Flight,ToF）与可见光果园苹果图像融合模型。对SPCNN模型引入带参数优化的双寻优策略,对非下采样轮廓波变换（Nonsubsampled Contourlet Transform, NSCT）融合规则进行改进。模型包括配准模块、编码区、多尺度分解模块、单目标SPCNN融合模型、多目标SPCNN融合模型、解码区。模型改进了SPCNN模型的参数优化方式以及迭代次数,模型自适应点火次数较低,在3～7次左右,具有点火次数低、自适应分割、效率高的优点。中光15:00时段点火识别成功率达到了100.00%,点火分割时间达到最低91.91s。与其他融合模型比较,模型在强光12:00、中光15:00、弱光18:20、19:00时段融合图像识别成功率达到100.00%;融合时间低于SPCNN模型,达到最低92.68s。模型识别精度最优达到了100.00%、融合耗时最低达到了92.68s、模型大小较SPCNN低一个数量级,可补充和完善图像层次融合理论和方法。  相似文献   

Molecular Characterization and Expression Profiles of Myrosinase Gene（RsMyr2） in Radish（Raphanus sativus L.）     

PAN Yan  ;XU Yuan-yuan  ;ZHU Xian-wen  ;LIU Zhe  ;GONG Yi-qin  ;XU Liang  ;GONG Mao-yong  ;LIU Li-wang 《中国农业科学(英文版)》2014,(9):1877-1888

Myrosinase is a defense-related enzyme and is capable of hydrolyzing glucosinolates into a variety of compounds, some of which are toxic to pathogens and herbivores. Many studies revealed that a number of important vegetables or oil crops contain the myrosinase-glucosinolate system. However, the related promoter and genomic DNA sequences as well as expression profiles of myrosinase gene remain largely unexplored in radish（Raphanus sativus）. In this study, the 2 798 bp genomic DNA sequence, designated as RsMyr2, was isolated and analyzed in radish. The RsMyr2 consisting of 12 exons and 11 introns reflected the common gene structure of myrosinases. Using the genomic DNA walking approach, the 5′-flanking region upstream of RsMyr2 with length of 1 711 bp was successfully isolated. PLACE and PlantCARE analyses revealed that this upstream region could be the promoter of RsMyr2, which contained several basic cis-regulatory elements including TATA-box, CAAT-box and regulatory motifs responsive to defense and stresses. Furthermore, recombinant pET-RsMyr2 protein separated by SDS-PAGE was identified as myrosinase with mass spectrometry. Real-time PCR analysis showed differential expression profiles of RsMyr2 in leaf, stem and root at different developmental stages（e.g., higher expression in leaf at cotyledon stage and lower in flesh root at mature stage）. Additionally, the RsMyr2 gene exhibited up-regulated expression when treated with abscisic acid（ABA）, methyl jasmonate（MeJA） and hydrogen peroxide（H2O2）, whereas it was down-regulated by wounding（WO） treatment. The findings indicated that the expression of RsMyr2 gene was differentially regulated by these stress treatments. These results could provide new insight into elucidating the molecular characterization and biological function of myrosinase in radish.  相似文献   

Variation of gynogenic ability in passion fruit (Passiflora edulis Sims.) accessions     

M. M. Rêgo  E. R. Rêgo  C. H. Bruckner  W. C. Otoni  C. M. Pedroza 《Plant Breeding》2011,130(1):86-91

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