CHARACTERIZATION AND MODE OF IN-VITRO REPLICATION OF PEA CHLOROPLAST ORIA SEQUENCES

被引:15
作者
REDDY, MK
CHOUDHURY, NR
KUMAR, D
MUKHERJEE, SK
TEWARI, KK
机构
[1] ICGEB,NEW DELHI 110067,INDIA
[2] JAWAHARLAL NEHRU UNIV,CBT,GENET ENGN UNIT,NEW DELHI,INDIA
[3] UNIV CALIF IRVINE,DEPT MOLEC BIOL & BIOCHEM,IRVINE,CA
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1994年 / 220卷 / 03期
关键词
D O I
10.1111/j.1432-1033.1994.tb18697.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A partially purified replicative system of pea chloroplast that replicates recombinant DNAs containing pea chloroplast origin sequences has been characterised. Polymerisation by this system is very fast and insensitive to chain terminators like dideoxynucleotides, arabinosylcytosine 5'-triphosphate, etc. Both strands of template DNA are synthesized and single-stranded DNA templates undergo more than one round of replication. When sequences of either of the two chloroplast origins of replication (OriA or OriB) are used as templates, the replicative intermediates are found to have sigma structures. Electron microscopic analysis of the sigma structures restricted with various enzymes reveals that the initiation site of in vitro replication maps near the displacement-loop regions where replication initiates also in vivo. Although the observed replication initiation in the OriA recombinant template is chloroplast-DNA-specific, the mode of replication is different from that observed in vivo with intact ctDNA. However, when the template DNA contains both the OriA and OriB sequences, the in vitro replication proceeds in the theta mode, the mode of replication usually observed in vivo.
引用
收藏
页码:933 / 941
页数:9
相关论文
共 31 条
[1]  
ALFANO C, 1989, J BIOL CHEM, V264, P10699
[2]  
BAKER TA, 1987, J BIOL CHEM, V262, P6877
[3]   A MODEL FOR INITIATION AT ORIGINS OF DNA-REPLICATION [J].
BRAMHILL, D ;
KORNBERG, A .
CELL, 1988, 54 (07) :915-918
[4]   CHLOROPLAST DNA-REPLICATION INVITRO - SITE-SPECIFIC INITIATION FROM PREFERRED TEMPLATES [J].
CARRILLO, N ;
BOGORAD, L .
NUCLEIC ACIDS RESEARCH, 1988, 16 (12) :5603-5620
[5]  
CHALLBERG MD, 1989, ANNU REV BIOCHEM, V58, P671
[6]   MINI-P1 PLASMID REPLICATION - THE AUTOREGULATION-SEQUESTRATION PARADOX [J].
CHATTORAJ, DK ;
MASON, RJ ;
WICKNER, SH .
CELL, 1988, 52 (04) :551-557
[7]   TRANSIENT FOREIGN GENE-EXPRESSION IN CHLOROPLASTS OF CULTURED TOBACCO CELLS AFTER BIOLISTIC DELIVERY OF CHLOROPLAST VECTORS [J].
DANIELL, H ;
VIVEKANANDA, J ;
NIELSEN, BL ;
YE, GN ;
TEWARI, KK ;
SANFORD, JC .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (01) :88-92
[8]  
DAVIS RW, 1971, METHOD ENZYMOL, V21, P413
[9]   CHLOROPLAST DNA DELETIONS ASSOCIATED WITH WHEAT PLANTS REGENERATED FROM POLLEN - POSSIBLE BASIS FOR MATERNAL INHERITANCE OF CHLOROPLASTS [J].
DAY, A ;
ELLIS, THN .
CELL, 1984, 39 (02) :359-368
[10]  
HSEIH CH, 1991, MOL GEN GENET, V225, P25