大豆查尔酮异构酶基因的克隆及乳酸菌表达载体的构建.pdfVIP

, Journ l ofAnhuiAgr.i Sci. 2010, 38( 19) : 9995- 9997 马树梅 况玲玲 * 刘洪禹, 王丕武 , 付永平, 张 卓, 马 超 (, 130118) [目的]将特异存在于豆科植物的 II型查尔酮异构酶 因CH I1A 构建到目前最有效的食品级乳酸乳球菌N ICE诱导表达系统中 [方法]利用RTPCR 技术从大豆总RNA 中克隆CH I1A 因,连入 pMD18T 克隆载体中并进行测序, 然后重组到乳酸乳球菌表达载体 PNZ8149CH I1A, 利用电穿孔方法转入乳酸乳球菌NZ3900中[结果]克隆得到CH I1A 完整开放阅读框670 bp, 与已报导序列 ( GenB nk AY595413)的同源性达到 99%; 通过 PCR和酶切鉴定, 成功地将CH I1A 导入到N ICE 表达系统中[结论]含有 CH I1A 因的乳酸乳球 菌高效诱导表达载体的构建为利用微生物发酵产生类黄酮奠定 础 大豆; 查尔酮异构酶 因; 乳酸乳球菌; N ICE系统 S 188 A 0517- 6611( 2010) 19- 09995- 03 C loning of Tap Chalcone Isomerases(Chi1a )Gene and Construction ofLactococcus L act is ExpressionV ector LIU Hongyu et al ( B iotechnolog ic l Center of Jilin Agr icu ltur lU niversity, Ch ngchun, Jinlin 130 118) A stract [ Objective] The mi w s to clone T pII ch lcone isom er se CH I1A from soybe n speci lly nd construct expression vector of PNZ8149CH I1A , nd then tr nsform it intoL ac tococcus lac tis N ICE systems. [M ethods] Ch lcone isom er se CH I1A w s cloned by RTPCR m ethod, nd it w s sequenced fter clonn ing into pMD18T vectors, nd recombined to expression vectorPNZ8149CH I1A , then it w s tr ns formed into L ac tococcus L actis NZ3900. [ R esults] T he sequencing results indic ted th t the cloned fr gm ent ofCH I1A cont ined 670 nucleo tides, nd sh red sequence homology of 92% w ith th t from Genb nk ccession numberAF595413 ( CH I1A ). CH I1A w s tr nsducted into N ICE expression systerm successfully by identific tion ofPCR nd digestion. [ Conclusion] The found tion of using them icroorg nism ferm en t tionm ethod to produce fl vono ids w s l id by construction of efficient induction expression vectorw ith ch lcone isomer se CH I1A. Key words Soybe n; Ch lcone isomer se; Lactococcus lac tis; The nisincontrol led expression ( fl vonoids) ( biofl conoids),