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1.
菌核青霉2246(Penicillium sclerotiorum)能够将人参皂苷Rg1转化为人参皂苷F1.以此菌为出发菌株,进行原生质体制备和再生的研究,确定原生质体的最佳形成条件:菌丝体培养24 h,用5 mg/mL溶壁酶、5mg/mL纤维素酶和5 mg/mL蜗牛酶的混合酶液进行酶解,以0.8 mol/L的KCI作为渗透压稳定剂,31℃水浴振摇2h.并对形成的原生质体进行亚硝基胍复合紫外线照射诱变,结果得到1株转化率显著提高、遗传性能稳定的诱变株( NU-1),其转化率由16.7%提高到30.5%.  相似文献   

2.
碱性普鲁兰酶产生菌的原生质体制备与诱变选育   总被引:1,自引:0,他引:1  
研究报道了Bacillus sp SX—12原生质体制备与再生最佳条件。实验表明,在液体完全培养基中加入2%的甘氨酸培养10h,原生质体制备最佳条件为:溶菌酶浓度0.5mg/mL,酶解温度37℃,酶解时间1.5h时原生质体形成率为93.8%。原生质体形成最佳高渗稳定剂为甘露醇,再生率26.4%。在原生质体制备的最佳条件下,用紫外线诱变技术选育产碱性普鲁兰酶的高产菌株。筛选到1株高产菌株SX—12C87,酶活由出发菌株的2.42μ/mL提高到6.87μ/mL,提高了约1.8倍。  相似文献   

3.
以紫杉醇产生菌树状多节孢HQD33的诱发突变株UL50-6和UL40-19 为出发菌株,将收集到的出发菌株UL50-6和UL40-19的菌丝体分别用pH5.5~6.0的0.7mol/L NaCl配制的3%溶壁酶、2%蜗牛酶、1%溶菌酶组成的复合酶系,30℃恒温酶解3~5h,制备原生质体。两菌株的原生质体经纯化后分别用热和紫外线灭活,其中UL50-6的原生质体在54℃热灭活5分钟,UL40-19的原生质体在30W紫外灯下,30cm,照射85秒进行紫外灭活,双亲株的原生质体存活再生率为零。同时对融合条件进行了初步探索,以含有Ca2 和Gly的35%~40%的PEG作为融合剂,融合时间为20分钟时,融合率可以达到4.44?0-2~6.92?0-2。对融合株TPF-1与双亲株的形态学、可溶性蛋白、过氧化物同工酶进行分析,确证其为双亲株的融合子。  相似文献   

4.
康氏木霉B—7和黑曲霉X—15原生质体的形成和再生   总被引:2,自引:0,他引:2  
研究了康氏木霉B-7和黑曲霉X-152株纤维素酶高产菌株的原生质体制备与再生。结果表明,采用纤维素酶、蜗牛酶、溶菌酶的混合酶液,可成功地制备2株真菌的原生质体。其中,B-7以这3种酶的配比6:5:2为最佳,X-15以8:4:2为最佳。原生质体形成的缓冲液系统均以0.2mol/L,pH6.0磷酸盐缓冲液为宜,渗透压稳定剂则分别以0.6mol/LNaCl和0.6mol/L蔗糖为宜。以菌龄18h(B-7)和16h(X-15)2株真菌的菌丝体,在37℃下酶解90min可获得最适量的原生质体,产量分别达9×106个/ml和1.9×107个/ml,且其再生率也较高,均达95%左右。  相似文献   

5.
影响枯草芽胞杆菌和荧光假单胞菌原生质体再生的因素   总被引:1,自引:0,他引:1  
目的:为了提高再生率,对影响革兰阳性菌枯草芽胞杆菌KR株和革兰阴性菌荧光假单胞菌B13株原生质体再生的因素进行研究。方法:研究了酶解时间,再生方式,再生培养基中稳定剂的种类,Ca^2+、Mg^2+、琥珀酸钠、L-色氨酸的浓度及培养基的放置时间对KR和B13株原生质体再生的影响。结果:对KR株酶解20min,采用夹层培养,再生培养基中加入0.6mol/L蔗糖、0.03mol/L Ca^2+、0.02mol/L Mg^2+、0.3mol/L琥珀酸钠、0.2mol/L L-色氨酸,培养基在37℃放置72h,原生质体再生率可达42.7%;对B13酶解15min,采用夹层培养,培养基中加入0.6mol/L NaCl、0.02mol/L Ca^2+、0.01mol/L Mg^2+、0.3mol/L琥珀酸钠、0.1mol/L L-色氨酸,培养基在37℃放置48h,原生质体再生率可达15.3%。结论:影响革兰阳性菌枯草芽胞杆菌KR株和革兰阴性菌荧光假单胞菌B13株原生质体再生的因素是不同的。  相似文献   

6.
以C.shehataeTZ8为出发茵株,利用1%溶壁酶和1%蜗牛酶酶解1.5h,制备成C.shehataeTZ8原生质体,并对原生质体进行紫外诱变,以含不同浓度乙醇的木糖液体培养基培养进行初筛和复筛,获得一株遗传性能稳定、耐乙醇能力达5.5%(v/v)的蕾株C.shehataeTZ8-4,比初始菌株耐乙醇能力提高了2%。对突变株C.shehataeTZ8-4发酵性能的研究结果表明:C.shehataeTZ8-4发酵糖能力从80g/L(葡糖糖和木糖比为2:1)提高到120g/L,最大乙醇产量从27.41g/L提高到43.12g/L。  相似文献   

7.
将 2株黑曲霉菌株 (I,II)在诱导产植酸酶的条件下分别制备成原生质体 ,并考察其原生质体形成及再生 ,经紫外诱变后 ,发现涂平板的黑曲霉 (II)原生质体在再生过程中菌落形态发生明显变化。考查突变菌种的植酸酶酶活 ,筛选到其中一个变异株比原始黑曲霉 (II)亲本株的植酸酶酶活提高了 2 .2倍。  相似文献   

8.
甘蓝型油菜与蔊菜的原生质体融合与植株再生   总被引:1,自引:0,他引:1  
以甘蓝型油菜下胚轴和蔊菜叶片为外植体提取原生质体, 采用PEG-高pH、高Ca2+附加DMSO的原生质体融合方法, 用液体浅层静置培养融合体, 获得了10株融合杂种, 观察了杂种形态学和细胞学。结果表明:1%纤维素酶+0.2%离析酶+3 mmol/L MES 酶解14 h 可获得较高产率的油菜原生质体, 0.25% 纤维素酶+0.5%离析酶+5 mmol/L MES酶解12 h可获得较高产率的蔊菜原生质体; 30% PEG + 0.3 mol/L葡萄糖+50 mmol/L CaCl2•2H2O +15%DMSO的融合条件下, 获得了10.4%的融合率; 实验所获的原生质体融合材料可作为新种质。  相似文献   

9.
毛栓菌原生质体制备和再生及单核菌株产漆酶特性   总被引:1,自引:0,他引:1  
毛栓菌Trametes hirsuta能有效地降解木质素,在生物燃料、制浆和饲料工业等方面具有很高应用价值。为了获得遗传性能稳定的T. hirsuta单核菌株,研究了其菌丝生长培养基的类型、菌丝生长时间(菌龄)、酶解时间、原生质体纯化离心速度和原生质体再生培养基类型对T. hirsuta YJ-9-1原生质体制备与再生的影响;采用DAPI染色和锁状联合缺失的观察,从再生株中筛选单核菌株并考察其产酶特性。结果表明:采用YGM菌丝生长培养基、88h菌龄、1h酶解时间、4,000r/min原生质体纯化离心速度以及YGMS再生培养基,最终可获得密度大约为5.0×106个/mL的原生质体悬浮液和9.1%的再生率;从200株再生菌株中筛选出了3株单核菌株,其中一株单核菌株D-2-1的漆酶产量比原菌T. hirsuta YJ-9-1明显提高,在第12天其漆酶酶活为771.67U/L,是原菌的1.51倍。  相似文献   

10.
紫外线诱变原生质体选育核黄素高产菌株   总被引:3,自引:0,他引:3  
以产核黄素生产用菌——阿舒假囊酵母RS-89为出发菌株,用对致期生长的菌丝体,经0.5%蜗牛酶+0.5%纤维素酶作用2h可获得大量原生质体,其再生率为6.1%。对经UV诱变的原生质体再生株进行初筛、发酵复筛,从中获得了菌落大、色素深、产量高于出发菌株30%的高产稳定株——UP-91。  相似文献   

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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.  相似文献   

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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.  相似文献   

17.
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.  相似文献   

18.
肝癌中HBV和HCV基因和抗原的分布及意义   总被引:1,自引:0,他引:1  
采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细  相似文献   

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For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.  相似文献   

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