[Cloning E. coli mRNAs with poly (A) tails by restriction digest polymerase chain reaction].
OBJECTIVE
To investigate the polyadenylation at the 3' terminal of the mRNAs in E.coli.


METHODS
mRNAs of E.coli was enriched from total RNA with oligo (dT)-cellulose, and reverse transcription was performed using oligo(dT)18 as primer prior to synthesis of double strands cDNA which was digested with Sau 3A I to produce multiple gene fragments that were then ligated with adapters. Restriction digest-polymerase chain reaction (RD-PCR) was employed to divide the fragments into 10 groups using 10 different combinations of the 4 primers, and the products were cloned into T-vectors.


RESULTS
More than 100 gene fragments were cloned, 30 of which were sequenced.


CONCLUSIONS
Polyadenylation of E.coli mRNA is not a biochemical curiosity, and very likely, it is a general attribute of the mRNAs of bacteria.
Hu
Zi-You
ZY
Institute of Molecular Biology, First Military Medical University, Guangzhou 510515, China.
Ma
Wen-Li
WL
Song
Yan-Bin
YB
Zhang
Bao
B
Wu
Qing-Hua
QH
Guo
Qiu-Ye
QY
Peng
Yi-Fei
YF
Zheng
Wen-Ling
WL
D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D008969:Molecular Sequence Data; D011061:Poly A / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D012333:RNA, Messenger; D011061:Poly A; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases