INFLUENCE OF ENZYME SOLUTION ON PROTOPLAST CULTURE AND TRANSIENT GENE-EXPRESSION IN MAIZE (ZEA-MAYS L)

被引：5

作者：

KRAUTWIG, B ^{[1
]}

LAZZERI, PA ^{[1
]}

LORZ, H ^{[1
]}

机构：

[1] AFRC, INST ARABLE CROPS RES, DEPT BIOCHEM & PHYSIOL, ROTHAMSTED EXPTL STN, HARPENDEN AL5 2JQ, HERTS, ENGLAND

来源：

PLANT CELL TISSUE AND ORGAN CULTURE | 1994年 / 39卷 / 01期

关键词：

BETA-GLUCURONIDASE; MACEROZYME; PLATING EFFICIENCY; PROTOPLAST YIELD;

D O I：

10.1007/BF00037590

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

The influence of two enzyme solutions, differing only in the presence or absence of Macerozyme, on protoplast yield, colony formation and transient GUS (beta-glucuronidase) activity was studied. For all parameters tested the presence of Macerozyme during protoplast isolation had a negative influence. Using an enzyme solution without Macerozyme suspension aggregates gave up to 4.4 times higher protoplast yield and plating efficiencies were increased up to 10-fold. Further, protoplasts isolated without Macerozyme showed a 5.2-fold higher GUS activity in transient gene expression. Apart from the presence of Macerozyme, longer incubation (3 compared with 1.5 h) of cell aggregates in the enzyme solution also had a negative effect on transient transformation efficiency. These data demonstrate that protoplast isolation conditions have a profound effect on transient gene expression and it is proposed that these factors will also influence stable transformation efficiency.

引用

页码：43 / 48

页数：6

共 21 条

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