用p389ARG7转化和诱变莱茵衣藻(C.reinhardtii)CBN1基因的研究  

Studies on Mutagenesis of CBN1 Gene of Chlamydomonas Reinhardtii by Transformation with Plasmid p389ARG7

作　　者：张文平[1] 徐琴[1] 杨红梅[1] 艾尔肯.热合曼 吾甫尔.米吉提 

机构地区：[1]新疆大学生命科学与技术学院,新疆乌鲁木齐830046

出　　处：《新疆大学学报（自然科学版）》2004年第3期288-291,299,共5页Journal of Xinjiang University(Natural Science Edition)

基　　金：国家自然科学基金 ( 3 0 0 60 0 3 5 ) ;新疆大学基金

摘　　要：用微玻璃珠研磨法将 p ARG7质粒转入到莱茵衣藻精氨酸依赖型、缺壁突变株品系 CC-1 61 8( arg7cw1 5 mt-)和 CC-1 61 7( arg7cw1 5 mt-)中 ,得到了一些不同的插入型突变转化子 .CC-1 61 7突变株品系的转化率为 72个 /μg DNA,CC-1 61 8突变株品系的转化率为 2 96个 /μg DNA,即转化率比 CC-1 61 7突变株品系高 .经多次转化共获得各种转化子 3 90 0多个 ,对它们进行荧光检测后检测出缺乏叶绿素 b的两个突变子ARGT3和 ARGT4,并进一步用高效液相色谱法对其光合色素含量进行检测结果证实 ,这两个突变体的确丧失了叶绿素 b合成能力 .它们将可作为The plasmid p389ARG7 is transformed into arginine dependent and cell wall-less strains CC-1618 (arg7cw15mt-) and CC-1617 (arg7cw15mt-) of Chlamydomonas reinhardtii by using glass bead method respectively, and some insertion mutated transformants with various background have been obtained. The transformation rate of the CC-1617 is 72 per μl DNA while that of the CC-1618 is 296 per μl DNA ,which shows that the transformation efficiency of latter is higher than that of the former. At last above 3 900 transformants have been obtained and fluorescence test is performed to screen them, from that 2 chlorphyll b-less mutants ARGT3 and ARGT4 have been detected and isolated . The further result of detection of photochrome contents screened by HPLC method demonstrates that each of the 2 mutants had lost the ability of biosynthesis chlorophyll b, and therefore they will become useful tool for the cloning and localization of the CBN1 gene.

关 键 词：莱茵衣藻 微玻璃珠研磨法 转化 叶绿素b基因 

分 类 号：Q942[生物学—植物学]