钝顶螺旋藻部分原生质体及单细胞的制备与培养

于1992年1月-1994年4月，进行超声波处理方法制备钝顶螺旋藻部分原生质体以作为基因工程受体，以及制备单细胞用于固体平板克隆化培养的研究。研究结果表明：超声波以20kHz频率、15W功率作用30s，可使藻丝体断裂成15．0±1．6个细胞长度；延长作用时间，至2－6个细胞长度时，细胞壁结构遭到破坏，形成部分原生质体；继续作用，可形成少量原生质体和大量单细胞。断裂藻丝体、部分原生质体、单细胞以及原生质体均可涂布于固体培养基上再生或生长。以一定密度涂布单细胞与原生质体，能够形成彼此分开的单个克隆，可用于筛选及遗传分析。本文提供了一种节省溶菌酶的制备螺旋藻透性体的方法，超声波作用利于外源基因的导入，而涂布培养利于进一步的筛选和形成克隆。

ISOLATION AND CULTURE OF PERMEAPLASTS AND SINGLE CELLS FROM SPIRULINA PLATENSIS

Jan., 1992 to April, 1994 studies carried out on isolation of spheroplasts forgene introduction and single cell for plating showed: 1. filaments of Spirulina platensis can be broken by ultrasonics into disired average length fragments; 2. whenthe length was 2-6 cells, cell walls were partially removed and permeaplasts wereformed; 3. with prolonged treatment few protoplasts and many single cells appeared;4. fragments single cells and protoplasts can regenerate or grow by plating on solidmedium; 5. plating single cells and protoplasts after dilution may be a useful method for cloning and selection. The results, suggesting a lysozyme-savlng method forpermeaplast preparation, indicate that ultrasonics is helpful for gene introduction,and that plating is good for genetic analysis and cloning of filamentous algae.