致病酵母菌基因组多态性及亲缘关系的研究

致病酵母是条件致病菌感染中最常见的菌群。其属间、种间及种内的分型具有重要的流行病学及临床意义。以随机扩增多态性(Randomly Amplified Polymorphism DNA markers,RAPD)的方法对48株临床上常见的酵母菌属间、种间及种内基因组型的多态性进行了研究,并以多种引物扩增带型的相似性系数的高低来评价酵母菌之间的亲缘关系。结果表明:RAPD带型可清楚的显示出假丝酵母(Candida)及相关酵母属间、种间及种内的差异,亲缘关系的研究表明假丝酵母属与隐球菌属(Cryptococcus)、丝孢酵母属(Trichosporon)的相似性系数为80％,除季也蒙假丝酵母(C. guilliermondii)外,假丝酵母属中不同种间的相似性系数为82％～87％,同种不同株间的相似性系数>90％。大多数属、种基因组分型的结果和形态学分类结果相符。     

用RAPD方法对平菇、香菇属间原生质体融合子的研究

RAPD是一种新的分子生物学技术。本实验用28个随机引物对平菇、香菇及其融合子进行PCR扩增, 在融合子谱带中,有64条与平菇相同(与平菇的相似指数SI=70.6%),14条与香菇相同(SI=33.6%),另外有26条谱带为双亲与融合子所共有。本实验结果说明融合子是双亲融合的产物,但在融合过程中,从双亲获得的遗传信息不等。本实验首次在食用真菌中获得了原生质体融合的分子生物学证据。我们认为,在食用菌研究中,RAPD将是一种有重要价值的分析方法。 Abstract:RAPD is a novel molecular biological technique.P.sapidus,L.edodes and their intergeneric fusants were tested by RAPD method using 28 randomly selected primers.In bands gained from fusants by amplification reaction,there were 64 bands with the same position as in P.sapidus (the similarity index (SI) was 70.6%),14 bands as in L.edodes.This results illustrated that fusants are true hybrid derived from protoplast fusion procedure,but the genetic information obtained from the two parents were not equal.Molecular biological evidence of protoplast fusion had obtained first time in edible mushroom by RAPD method in this experiment,we suggest that this technique is a valuable genetic marker for mushroom study.     

用RAPD方法对平菇、香菇属间原生质体融合子的研究

RAPD是一种新的分子生物学技术。本实验用28个随机引物对平菇、香菇及其融合子进行PCR扩增, 在融合子谱带中,有64条与平菇相同(与平菇的相似指数SI=70.6%),14条与香菇相同(SI=33.6%),另外有26条谱带为双亲与融合子所共有。本实验结果说明融合子是双亲融合的产物,但在融合过程中,从双亲获得的遗传信息不等。本实验首次在食用真菌中获得了原生质体融合的分子生物学证据。我们认为,在食用菌研究中,RAPD将是一种有重要价值的分析方法。 Abstract:RAPD is a novel molecular biological technique.P.sapidus,L.edodes and their intergeneric fusants were tested by RAPD method using 28 randomly selected primers.In bands gained from fusants by amplification reaction,there were 64 bands with the same position as in P.sapidus (the similarity index (SI) was 70.6%),14 bands as in L.edodes.This results illustrated that fusants are true hybrid derived from protoplast fusion procedure,but the genetic information obtained from the two parents were not equal.Molecular biological evidence of protoplast fusion had obtained first time in edible mushroom by RAPD method in this experiment,we suggest that this technique is a valuable genetic marker for mushroom study.     

PCR技术检测人乳头状瘤病毒DNA

本文用一对适合人乳头状瘤病毒DNA的通用PCR引物对临床阳性和正常组织进行了PCR扩增检测分析,阳性组织均得到一条特异性的DNA扩增片段,正常组织均没有任何扩增片段。阳性组织DNA扩增片段经克隆后进行DNA序列分析,证明该扩增片段确为目标DNA扩增片段。     

氨基载体共价结合固定化海洋假丝酵母脂肪酶

共价结合法是重要的工业酶固定化方法,利用稳定的共价键固定化工业酶,在载体和酶间形成多点共价连接,可以制备稳定性较好的固定化酶,更具有实际应用价值。利用氨基载体共价结合固定化海洋假丝酵母脂肪酶,采用较为廉价的戊二醛进行辅助交联,通过单因素和正交试验,确定最佳固定化条件为:25℃、pH5. 0、0. 1%戊二醛、0. 25g载体、交联0. 5h、固定化1h、加酶量为800U,最终得到的固定化酶酶活达到83. 01U/g。固定化脂肪酶的最适pH较游离酶向碱性方向偏移,最适反应温度提高10℃,固定化酶的热稳定性和酸碱稳定性比游离酶好且重复使用性和储存稳定性明显优于游离酶。同时发现交联剂是制备固定化脂肪酶的重要因素,因此探索新型交联剂对于固定化效果的提高具有重要意义,为海洋假丝酵母脂肪酶的固定化工艺技术和工业应用奠定了良好基础。