CENTENNIAL HONORS COLLEGE
Western Illinois University
Undergraduate Research Day 2016
PosterPresentation
EnhancedGeneTransferinLeuconostoc
AllisonHebert
FacultyMentor:ScottHolt
Biology
ThegoalofthisstudyistoimproveefficiencyoftransformationinLeuconostoc-anon-pathogenicgrampositive bacterial genus that makes natural products including complex sugars derived from sucrose.
Complexsugarshavemanyapplications,e.g.,prebioticsandcosmetics.Thesecomplexsugarsaremade
fromanenzymecalledsucrase.Leuconostocexcretessucrasewhengrowninthepresenceofsucrose.
ThemechanismbywhichLeuconostocproducessucraseisunknownduetolackofgenetictoolsneeded
forstudyingtheprocess.Todiscoverthemechanismsbehindsucraseproductionandenhancefurther
research and industrial utilization of the produced complex sugars, an efficient method of
transformationmustbedevised.
LeuconostocdoesnotacceptforeignDNAeasily,mostlikelyduetoitsthickcellwall.ForLeuconostocto
acceptforeignDNAamethodcalledelectroporationmustbeused.Thismakesitpossiblefortheforeign
DNAtoenterLeuconostoccellsbutalsoweakensthem.Manydonotsurvive.Thisstudywillattemptto
increasethenumberofcellsthatsurviveandtheefficiencyoftransformationbytreatingtheweakened
cellsafterelectroporationduringaresuscitationstep.Improvementofthetransformationefficiencyof
Leuconostoc will be measured by comparing control cells to experimental cells treated with either
Oxyrase® or an osmotic stabilizer called sorbitol. The resuscitated Leuconostoc will be grown on a
nutrient agar containing chloramphenicol. If the resuscitated Leuconostoc cells grow on the nutrient
agarcontainingtheantibioticitwillindicatesuccessfultransferoftheplasmid.