He-Ne激光在异种间原生质体融合中的应用

用632.8nm 12mW的氦氖激光及聚乙二醇（PEG）复合诱导融合枯草芽孢杆菌与黑曲霉原生质体.为了提高枯草芽孢杆菌的糖化酶产酶能力,对两亲本在融合中采用不同的激光照射时间,选育出的融合子与枯草芽孢杆菌亲本相比,糖化酶酶活提高了2倍,并通过酯酶同工酶谱分析对融合子进行了鉴定,结果表明:融合子遗传性状与亲本相比发生了显著变化,通过传代培养,融合子具有良好的遗传稳定性.     

He-Ne激光诱变黑曲霉Sx的原生质体

对生淀粉糖化菌黑曲霉Sx用混和酶制取原生质体进行了探索.在GM培养基中加入一定量的Cu²⁺、Mn²⁺,原生质体产量较高;采用pH4～5的蜗牛酶、纤维素酶、溶菌酶的混合酶,最佳浓度比为:1.5:3.5:1.5(mg/mL).取在GM培养基中培养17～18h的菌丝,在32℃酶解3h,原生质体产量最高,达到2.4×10⁵个/mL;在加入β-巯基乙醇及二硫苏糖醇处理菌丝时,原生质体产量提高近3倍.在再生培养基中,加入终浓度为30mmol/LMgSO₄时,原生质体再生率由16.7%提高到25.6%.用He-Ne激光照射原生质体时,功率9mW照射30min时致死率为50%,随着时间的延长,原生质体存活率降低.采用波长632.5nm、功率9mW、光斑直径5mm的He-Ne激光多次照射诱变,筛选出一株酶活力最高菌株黑曲霉Sy,活力提高51%,经连续传代5次,酶活力稳定.     

氦-氖激光诱变细菌细胞及原生质体选育果胶酶高产菌株

利用氦-氖激光(波长632.8nm,功率15mW)诱变果胶酶产生菌的菌体细胞,获得了突变株ZH-g,其酶活达到301 u·mL－1,是出发菌株的1.3倍.进一步用氦-氖激光照射ZH-g的原生质体,从而得到了高产突变株ZH-gA,其酶活达到357 u·mL－1,比ZH-g提高了18%,是出发菌株的1.6倍.该菌株在好氧和静置条件下均能良好产酶,经传代实验证明其产酶性能稳定.     

He-Ne激光诱变原生质体选育四环素高产菌的研究

采用He-Ne激光对金霉素链霉菌(S.aureofaciens)原生质体诱变处理,结果表明:He-Ne激光对该菌原生质体具有明显的致死作用.经激光处理的原生质体再生株产抗生素能力与亲本菌株相比,四环素产量平均提高9.32%,最高提高20.6%.     

金盏菊原生质体对土壤铅/镉响应机制的FTIR,2D-IR和XPS证据

土壤修复是"十四五"期间国家重点支持的环保领域,是实现社会可持续发展的重要保障.与其他方法相比,植物修复技术整体优势突出,对于土壤重金属的去除净化更为有效.原生质体是植物细胞代谢活动的重要场所,相对于细胞壁而言,原生质体对重金属胁迫的生理响应同样强烈.现阶段,同类植物修复机制研究多从分子生物学层面切入;本研究则从谱学角...     

氦氖激光对链霉菌原生质体种间融合频率的影响

采用双亲灭活原生质体融合技术对金霉素链霉菌(Streptomyces aureofaciens)和龟裂链霉菌(Streptomyces rimous)种间原生质体融合进行了研究,分别利用聚乙二醇(PEG)、He-Ne激光以及PEG与He Ne激光混合诱导融合,结果表明:PEG及激光的双重诱导作用,使链霉菌原生质体种间融合频率得到显着提高。通过同工酶谱分析对融合子进行了初步鉴定。     

He-Ne激光对γ-亚麻酸产生菌少根根霉的诱变作用

采用He-Ne激光和化学诱变剂LiCl对少根根霉孢子和原生质体进行复合诱变,从一株产GLA的出发菌株少根根霉R8#中选育出高产菌株RC378.摇瓶培养含油脂量47.8%,GLA含量12.6%,比出发菌株油脂含量提高了130.8%,GLA含量提高了276.7%.从菌落形态、同工酶酶谱、红外光谱吸收特征和油脂产量及组成比较分析表明:该菌株较出发菌株具有显著的变异.菌株RC378经传代培养,产脂量维持在41.1%～48.7%,GLA含量维持在10.8～12.6%.该菌株具有较好的遗传稳定性.     

Visualized investigation of yeast transformation induced with Li+ and polyethylene glycol

The effects of Li⁺ and polyethylene glycol (PEG) on the genetic transformation of Saccharomyces cerevisiae were investigated by using fluorescence microscopy (FM) to visualize the binding of plasmid DNA labeled with YOYO-1 to the surface of yeast cells, scanning electron microscopy (SEM) and atomic force microscopy (AFM) to image the change in surface topography of yeast cells, coupled with transformation frequency experiments. The results showed that under the same conditions, the transformation frequencies of yeast protoplasts were much higher than those of intact yeast cells. PEG was absolutely required for the binding of DNA to the surface of intact yeast cells or yeast protoplasts, and had no effect on the surface topography of intact yeast cells or yeast protoplasts. In the presence of PEG, Li⁺ could greatly enhance the binding of plasmid DNA to the surface of intact yeast cells, increase their transformation frequency, and affect their surface topography. On the other hand, no effect on the DNA binding to the surface of protoplasts and no increase in the number of transformants and no surface topography changes were found upon the treatment with Li⁺ to protoplasts. In the present work, the effects of Li⁺ and PEG on yeast genetic transformation were directly visualized, rather than those deduced from the results of transformation frequencies. These results indicate that cell wall might be a barrier for the uptake of plasmid DNA. Li⁺ could increase the permeability of yeast cell wall, then increase the exposed sites of DNA binding on intact yeast cells. The main role of PEG was to induce DNA binding to cell surface.     

氦氖激光对链霉菌原生质体种间融合频率的影响

采用双亲灭活原生质体融合技术对金霉素链霉菌（Ｓｔｒｅｐｔｏｍｙｃｅｓａｕｒｅｏｆａｃｉｅｎｓ）和龟裂链霉菌（Ｓｔｒｅｐｔｏｍｙｃｅｓｒｉｍｏｕｓ）种间原生质体融合进行了研究，分别利用聚乙二醇（ＰＥＧ）、ＨｅＮｅ激光以及ＰＥＧ与ＨｅＮｅ激光混合诱导融合，结果表明：ＰＥＧ及激光的双重诱导作用，使链霉菌原生质体种间融合频率得到显著提高．通过同工酶谱分析对融合子进行了初步鉴定．