[NIES-90 microcystin producing algae strain genome library construction and one isolated gene analysis].
OBJECTIVE
To construct a plasmid microcystis genome library for microcystin (MC) pertaining gene family screening or microcystis strain specific gene screening.


METHODS
Extracting genome DNA from MC producing strain NIES-90, then digesting DNA by HindIII and ligating to CIPase treated pUC18/HindIII fragment which later transformed to JM109. After checking the library by Xgal/Antibiotics plate, clones isolation, a few clones were sequenced and specific primers synthesized; finally, PCR was used to testify clone's NIES-90 strain specificity.


RESULTS
Successfully constructed a NIES-90 genome plasmid library with following functions: a) cloning efficiency more than 95% by comparing the white/blue clones ratio in Xgal Plate; b) 200-700 bp average insert length with 3 x 10(5) independent clones by independent clones analysis; c) 10 times more coverage of original NIES-90 genome; d) strong strain specificity by cross PCR analysis of some clone's sequence in two different microcystis strains compared with FACHB-469.


CONCLUSIONS
We successfully constructed a NIES-90 MC producing microcystis strain's genome plasmid library which could be used for screening MC pertaining gene family because of high strain specificity of some clones.
Zhou
X
X
Institute of Preventive Medicine, Fudan University, Shanghai 200032, China.
Yao
L
L
Yu
S
S
D018744:DNA, Plant / Q000032:analysis; D056890:Eukaryota / Q000235:genetics*; D015698:Genomic Library; D052998:Microcystins; D010456:Peptides, Cyclic / Q000235:genetics*; D016133:Polymerase Chain Reaction
D018744:DNA, Plant; D052998:Microcystins; D010456:Peptides, Cyclic; C078588:microcystin