31219.人類未知功能蛋白質(zhì)的克隆、表達(dá)、純化和結(jié)晶

1、東北師范大學(xué)碩士學(xué)位論文人類未知功能蛋白質(zhì)的克隆、表達(dá)、純化和結(jié)晶姓名：韓振坤申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：細(xì)胞生物學(xué)指導(dǎo)教師：陸軍鄭曉峰20090501AbstractFollowingthecompletionofthehumangenomesequence，muchattentionisnowshiftingtowardsthefunctionsofthegeneproductsThehie)putthroughcrystallizat

2、ionanddeterminationofunknownfunctionproteinwasamainapproachestoexplaintheirfunction100humangeneswereselected邪testtargetsforparallelcloning，expression，purificationandcrystallizationProteinsfromthesegeneswereselectedtohave

3、amolecularweightofbetween20and50kDanottohaveahighpercentageofhydrophobicresidues(iemorelikelytobesoluble)andtohavenoknowncrystalstructuresandwerenotknowntobesubunitsofheterocomplexesTodate，100expressioncloneshavebeencons

4、tructedwiththeGatew礦cloningsystemOfthese，79cloneswereexpressedasrecombinantproteinsinEscherichiacolistrainBL21(DE3)，and9cloneswereexpressedasrecombinantproteinsinEscherichiacolistrainRosetta(DE3)，ofwhich21weresolubleand1

5、2havebeenpuriedtohomogeneityCrystallizationconditionswerescreenedforthepurifiedproteinsin48一wellplatesbythesittingdropAfterfurtherrefmementwiththesamedeviceorbythehangingdropmethod，ThecrystalofCOQ3weregrownTheresultsprov