Cloning, molecular characterization and expression analysis of two small heat shock genes isolated from wine Lactobacillus plantarum.
OBJECTIVE
Understanding the molecular response to stress tolerance of wine Lactobacillus plantarum.


RESULTS
Two genes codifying for heat shock proteins were cloned from wine L. plantarum. The coding regions of the two heat shock genes are 420 and 444 nucleotides long, and started with an ATG codon suggesting that they were translated. The protein sequences deduced from the isolated genes have a molecular mass of 18.483 and 19.282 kDa, respectively, and were therefore named hsp18.5 and hsp19.3. The expression of small heat shock genes was analysed by RT-PCR analysis. Moreover, the 5' and 3' noncoding regions were cloned and sequenced.


CONCLUSIONS
The expression of the heat shock genes was strongly induced by heat, cold and ethanol stress. Analysis of the 5' and 3' flanking regions of hsp18.5 and hsp19.3 genes, revealed the presence of an inverted repeat sequence (TTAGCACTC-N(9)-GAGTGCTAA) homologue to the CIRCE elements found to the upstream regulatory region of heat shock operons, and an inverted sequence that could form a stem and loop structure that it is likely to function as a transcriptional terminator. Based on their structures, the genes were classified as belonging to Class I of heat shock genes according to the B. subtilis nomenclature of heat response genes.


CONCLUSIONS
Small heat shock genes isolated from wine L. plantarum might have a role in preventing damage by cold stress.
Spano
G
G
Department of Food Science, Foggia University, Foggia, Italy. g.spano@unifg.it
Capozzi
V
V
Vernile
A
A
Massa
S
S
D000595:Amino Acid Sequence; D003001:Cloning, Molecular / Q000379:methods; D003062:Codon / Q000235:genetics; D003080:Cold Temperature; D000431:Ethanol / Q000378:metabolism; D015870:Gene Expression; D005798:Genes, Bacterial / Q000235:genetics*; D006360:Heat-Shock Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D006358:Hot Temperature; D048191:Lactiplantibacillus plantarum / Q000235:genetics* / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014920:Wine / Q000382:microbiology*
D003062:Codon; D006360:Heat-Shock Proteins; D000431:Ethanol