有淀粉糖化酶活性的耐高温酿酒酵母的构建研究

通过诱导Hu—TY一1(耐高温酿酒酵母)产孢及单孢分离得到的单倍体株Hu—TY—l—A(Sa-ccharamyces cerevisiae)与具有STAl、STA2、STA3(淀粉糖化酶)遗传因子的酵母(S．Dias-taticus)单倍体和纯合二倍体进行相同交配型的种间原生质体融合,融合率为3．3×1O^-6-8．7×1O^-6。融合子的细胞形态、大小、DNA含量、生长和发酵特性等方面均不同于亲株。融合株BA-2、BA-3、CA-18、DA-2、CCA-30、其细胞体积与DNA含量为亲株之和,30℃生长速率与生物量为亲株的2—3倍,40℃生长速率与生物量为亲株HU－TY－1－A的1．3－1．6倍,能够利用淀粉,并且在40℃高温葡萄糖发酵酒精产率比糖化酵母高,比HU－TY－1－A低,是一些有淀粉糖化酶、耐高温的酿酒酵母新菌株。     

酿酒酵母与糖化酵母的种间原生质体融合及其融合子的鉴定

本文报道了酒精生产菌株K氏酿酒酵母Sacchormyces cerevisiae var.ellipsoideus的HUK-1(his-,二倍体,但在我们所试的5种产孢培养基上均不产孢)与糖化酵母Sac—charomyces diastaticus 7c(arg-,a)的种间原生质体融合,其营养标记互补的融合频率约是2．07×10^-6—3．40×10^-5。这些融合子曾在选择培养基MMs或Mmo上连续传代10次,以促进两亲核的融合。融合子的酒精发酵特性,细胞形态、体积大小、DNA含量、繁殖速率、发酵强度以及产孢能力等方面的观察和测定结果表明,均不同于双亲菌株。用显微操作器解剖了个别原养型融台子HU—KDF—185的4孢子子囊,在获得的93个单孢株中,其淀粉发酵特性和遗传标记均有双亲类型的分离或重组现象。上述实验结果充分证明了不同倍性的酵母之间可以通过原生质体融合获得种间杂种。     

酿酒酵母和产朊假丝酵母原生质体的形成、再生及属间融合

使用由亚硝基胍诱变所得到的营养缺陷型作为单倍体融合亲株的核基因标记,同时也采用线粒体球红霉素抗性突变株的小菌落形式作为融合亲株的线粒体基因标记。酿酒酵母(Saccharomyces cerevisiae)和产朊假丝酵母(Candida utilis)两亲株原生质体的制备是用对数生长早期的细胞在蜗牛酶和0.7M KCl及β-巯基乙醇或二巯基苏糖醇的作用下完成的。二者的原生质体的形成率在30—60分钟内达到90—99％。原生质体再生率,酿酒酵母最高为29—35％,产朊假丝酵母为7.5％。两亲株的原生质体在35％PEG(M.W.6,000),10mM CaCl_2条件下被诱导融合。在基础培养基上,长出以营养互补为标记的融合菌株。融合频率为10~(-5)—10~(-6)。试验表明,这些融合菌株具有杂种的性质。其中一株杂合子在同化D-木糖、纤维二糖等的能力上比亲株明显增强。     

酿酒酵母原生质体融合的研究

本实验通过酿酒酵母(Saccharomyces cerevisiae)营养缺陷型菌株H802-9A(a ade~-his~7 cys~2 tyr~1)和H802-3 A(a ade~- cra~1)进行原生质体融合,获得营养互补的重组融合子,选择两亲株的对数生长早期细胞(2-4×10~7个/m1),在0.1％β-琉基乙醇及1-2％蜗牛酶的作用下,原生质体形成率达99-100％,在35％聚乙二醇(MW6000)-10mMCaC1溶液的作用下,可获得融合率为2.6×10~(-6)-2.1×10~(-7)的杂合子,自发回复突变率1.5-7×10~(-9)。     

电诱导酵母属与短梗霉属属间融合的研究

本文报道了经GH一401 型电诱导细胞融合、基因转移仪诱导实现的酿酒酵母(Saccharomyces cerevisiae NK491) 与出芽短梗霉 (Aureobasidium pullulans NKB93—0061eu)的原生质体电诱导融合。在融合小罐中用3个强度为11 Kv／cm,时程为10μs的高压电脉冲处理原生质体,获得的融合子营养互补的融合频率为5.8×10^-5。在选择培养基上连续传代10次,然后在完全培养基上传代20次,最终得到4株稳定的融台子。融合子的细胞形态、大小、核的DNA含量以及酒精发酵等特性的观察和测定表明,4株融合子与双亲均有明显差异。实验结果表明,电诱导原生质体融台对酵母菌的属间融合是一种行之有效的方法,具有较高的融合频率。     

部分酶解酵母高效电击转化研究

以酵母质粒YCp50为外源DNA,电击转化部分酶解酵母宿主菌AB1380,转化效率稳定在10~6转化子/μg质粒DNA左右,比不酶解酵母或酵母原生质球作受体的电击转化效率高一个数量级以上,也比PEG介导的酵母原生质球转化高3～5倍,而且适合于大片段DNA如水稻YAC分子的转化。达最佳转化时的有关技术参数为:新接菌种通气培养至细胞密度1×10~8～1.5×10~8个/ml;转化时细胞密度控制在1×10~9～1.5×10~9个/ml;每毫升酶解缓冲液加15u溶菌酶(lyticase),30℃下处理酵母5min进行部分酶解;电击时,电场设置在6.25kV/cm、电容25μF,电击后直接铺板。     

酿酒酵母和粟酒裂殖酵母属间融合和融合子特性

酿酒酵母(Saccharomyces cerevisiae Hansen)PW218和粟酒裂殖酵母(Schizosaccharomyces pombe Lindn)PW232的原生质体用20mmol/L CaCl_2和30％PEG(MW6000)处理进行属间融合,获得了10多株融合子,融合率为0.65～1.96×10~(-5)。对F_2和F_(10)两株融合子进行了葡萄糖、木糖及葡萄糖和木糖混合液的摇瓶实验结果表明F_(10)融合子利用葡萄糖、木糖及两种糖混合液产乙醇的能力大大高于两亲株。F_2融合子对木糖以及葡萄糖和木糖混合液的发酵能力亦较两亲株高,其中利用木糖产乙醇的量分别比PW218和PW232提高1.38倍和2.65倍。     

H~+、N~+、Ar~+注入对啤酒酵母存活率的影响及SEM和ESR研究

研究了H+ 、N+ 、Ar+ 低能离子注入酵母菌剂量对存活率的影响 ;离子注入对酵母细胞的刻蚀损伤作用和离子注入后对细胞内自由基产生的影响。结果表明 :酵母的存活率随着注入剂量的增加而减少 ,离子注入对其存活率影响程度是H+ >N+ >Ar+ ,其半致死剂量分别是 2 .1× 10 1 4 ions cm2 ,5 .5× 10 1 4 ions cm2 ,6 .8× 10 1 4 ions cm2 。随着离子注入剂量的增加对酵母细胞的损伤和刻蚀作用逐渐增大 ,刻蚀损伤作用具有不均匀性 ;离子注入后酵母细胞内自由基产额明显增加 ,随着注入剂量的增大 ,自由基的强度也增大 ,逐渐呈饱和趋势     

电融合技术选育能利用木糖和纤维二糖生产乙醇的菌株

以携带营养缺陷型标记的季也蒙假丝酵母(Candida guilliermondii s 208 Arg-)和酿酒酵母(Saccharomyces cerevisiae 314)为亲本,采用GH-40l型电诱导基因转移／细胞融合仪,用3个强度为18kV／cm,时程为10μs,间隔为1 s的高压电脉冲处理原生质体,营养互补的融合频率为3．6×10^-3。在选择和非选择培养基上连续传代20余次后,对得到的稳定的融合子进行分析比较,结果表明,它们是两亲株融合后形成的融台体。其中有两株融合子能利用木糖和纤维二糖生产乙醇,F－106利用D－木糖(2％)和纤维二糖(2％)生产乙醇的产量分别为3．1g／l和1．05g／l,F－308分别为0．2g／I和2．8g／l。     

酵母融合菌株的构建及其特性研究

以酿酒酵母和糖化酵母为亲本,通过正交实验优化了亲本原生质体的制备,再生以及融合的最佳条件,由此获得了既具有糖化能力又可产高浓度酒精的稳定的酵母融合菌株。融合株的体积较亲株大,ＤＮＡ含量与两亲株ＤＮＡ含量之和相当,淀粉水解能力,葡萄糖发酵效率,生长速率及产酒精能力等均优于亲株     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.