排序方式: 共有40条查询结果,搜索用时 250 毫秒
1.
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
2.
3.
4.
BACKGROUND: The aims of the present study were to validate a vital mitochondrial potentiometric staining method in Chlamydomonas reinhardtii and to utilise this method to examine the effect of the herbicide bromoxynil octanoate on mitochondrial potential in this species. A range of stains was investigated, including Rhodamine 123, DASPMI, Mitotracker Green, Mitotracker Orange and JC‐1. RESULTS: Rhodamine 123 (R123) had the highest utility of several candidate stains. Incubation with both 5 and 10 µM carbonyl cyanide 3‐chlorophenylhydrazone caused significant fluorescence collapse [Dunn's post test (40.00, P < 0.01) and (45.49, P < 0.01) respectively], demonstrating that the R123 fluorescence reported mitochondrial potential. The effect of the herbicide bromoxynil octanoate was examined. Exposure to 0.1 mM of bromoxynil resulted in a significant increased mitochondrial fluorescence compared with the baseline (Mann–Whitney U = 222, P < 0.002), while concentrations of 1 mM and greater resulted in significant, almost complete loss of mitochondrial potential [mean fluorescence ratio = 1.193–1.289 (where a ratio of 1 represents total potential loss), Mann–Whitney U = 0.0, P < 0.001 (1 mM ), 0.0, P < 0.0001 (2 mM ), 0.0, P < 0.0001 (5 mM )]. EC50 of the collapse in mitochondrial potential owing to bromoxynil incubation occurred at 0.72 mM , and the mean t50 of bromoxynil octanoate action was 93 s. CONCLUSIONS: R123 is a sensitive potentiometric dye in C. reinhardtii that may find further use in investigations of both mitochondrial bioenergetics in plants and environmental toxicology. Copyright © 2011 Society of Chemical Industry 相似文献
5.
缺硫对莱茵衣藻叶绿素荧光参数和产H2速率的影响 总被引:2,自引:0,他引:2
采用TAP及TAP-S培养基培养莱茵衣藻(Chlamydomonas reinhardtii D.),测定了该藻叶绿素荧光参数及产氢气能力.结果表明:莱茵衣藻在TAP培养基内生长良好,有微量氢气产生,最高产氢速率只有1.4×10-4 ml/(mgChl·h).在TAP-S培养基内,莱茵衣藻的荧光参数Fv/Fo、ΦPSⅡ、qp分别在40~48 h下降到初始值的50%,说明缺硫对藻光合作用的影响首先发生在天线色素到PSⅡ反应中心的传能过程以及光合作用暗反应所需的酶.TAP-S培养基内藻最高产氢速率达0.22ml/(mgChl·h),缺硫可以显著提高莱茵衣藻氢气产生的速率. 相似文献
6.
本文研究了不同pH(5.0、7.0和9.0)和F-浓度(0.1、1、10、50、100、200mmol·L-1)对莱茵衣藻和蛋白核小球藻的胞外碳酸酐酶活性、PSⅡ实际光合效率ΦPSⅡ和叶绿素a合成量的共同影响。结果表明:在低于1mmol·L-1的F-作用下,两种微藻的生理和生长指标主要受pH影响;酸性条件下,两种微藻胞外碳酸酐酶活性显著低于中性和碱性条件时,ΦPSⅡ随pH升高而增大,叶绿素a合成量随pH升高而增加。在高于1mmol·L-1的F-作用下,两种微藻的生理及生长指标受F-和pH共同影响,且F-的作用大于pH的作用,胞外碳酸酐酶活性随着F-浓度升高先增加后降低,同时随着pH下降而降低;ΦPSⅡ和叶绿素a合成量则随着F-浓度升高和pH下降而迅速下降。胞外碳酸酐酶活性、ΦPSⅡ和叶绿素a合成量都能够作为指示微藻受氟胁迫的指标,以微藻为材料除去自然界氟超标水体中的超标F-,理论上是可行的。 相似文献
7.
微藻用于处理养猪场废水并同时生产可再生能源微藻生物质,由此实现养猪场废水的生物资源化修复。同时利用2种不同微藻即小球藻和莱茵衣藻,处于不同温度环境(15 ℃或25 ℃)条件下,对具有不同起始氨氮浓度(NH4+-N 浓度分别为100 mgL-1或250 mgL-1)的养猪场废水处理效果观察,分别测定处理后养猪场废水中COD去除能力和微藻生物质累积能力。结果显示,小球藻和莱茵衣藻混合微藻在较高的养猪场废水起始氨氮浓度和较适宜生长条件下(25 ℃)具有较高效率的废水处理能力和生物质累积能力。 相似文献
8.
人白细胞介素4在衣藻叶绿体中的高效转化及表达 总被引:3,自引:1,他引:3
人白细胞介素4(hIL4)是一种重要的免疫活性调节分子,与一些自身免疫性疾病有重要关系。将hIL4基因融合到衣藻叶绿体rbcL 5-’UTR和3’-UTR之间,组成5’UTR-hIL4-3’UTR片段,将其插入到来源于衣藻叶绿体基因组5.7 kb同源片段中,构建成pXhIL4同源重组质粒。利用电脉冲法将该质粒与含有aadA基因的壮观霉素抗性质粒p228共同转化衣藻叶绿体,从含100μg/mL的壮观霉素抗性平皿上筛选到阳性藻落,并通过PCR方法检测到hIL4基因,其共转化频率高达90%。经过western blotting印迹转移分析,证实了人白细胞介素4基因在衣藻叶绿体中获得成功表达,为进一步研究在衣藻叶绿体中高效表达外源蛋白奠定了基础。 相似文献
9.
莱茵衣藻磷酸果糖激酶RNAi载体构建
及其对莱茵衣藻油脂积累的影响 总被引:3,自引:0,他引:3
为研究磷酸果糖激酶(PFK2)对微藻油脂积累的影响、克隆了莱茵衣藻磷酸果糖激酶同源基因CrPFK2、
构建CrPFK2 RNAi 干涉载体并转化莱茵衣藻。通过测定转基因藻在HSM 培养下的生物量和油脂含量的结果发现、
CrPFK2 RNAi 转基因藻株在HSM 培养基中生长加快、油脂含量下降了17.03%耀21.48%、说明CrPFK2 基因表达的降
低与藻细胞油脂含量减少成正相关。CrPFK2 通过改变光合碳流的多少间接调控藻细胞油脂的合成。该结果为
CrPFK2 基因应用于微藻油脂的遗传改良将起到重要作用。 相似文献
10.
研究了不同饥饿时间下大型溞Daphnia magna (体长约3 mm)对沙角衣藻Chlamydomonas sajao和蛋白核小球藻Chlorella pyrenoidos的摄食选择性。沙角衣藻与蛋白核小球藻相比,具有较高的适口性,但营养价值相对较低。按照沙角衣藻和蛋白核小球藻的比例分别为5×104.35×104、20×104.20×104、35×104.5×104 cells/mL混合投喂饥饿0、0·25、1、4、8 d的大型溞,3 h后测定大型溞的摄食率、滤水率和食物选择系数。结果表明:与饱食组和短时间饥饿(0·25、1 d)组相比,长时间饥饿(4、8 d)导致大型溞对蛋白核小球藻的摄食率和滤水率显著增加(P<0·05),而对沙角衣藻的摄食率和滤水率显著降低(P<0·05);饱食和短时间饥饿状态下大型溞优先摄食沙角衣藻,而在长时间饥饿状态下大型溞优先摄食蛋白核小球藻;大型溞在长时间饥饿和短时间饥饿状态(含饱食)下的最优觅食行为不受食物中沙角衣藻与蛋白核小球藻相对密度的影响。研究表明,大型溞的饥饿状态可以影响其对藻类食物的选择性。 相似文献