福建师范大学学报(自然科学版)
福建師範大學學報(自然科學版)
복건사범대학학보(자연과학판)
JOURNAL OF FUJIAN TEACHERS UNIVERSITY（NATURAL SCIENCE）
2010年
1期
96-99
,共4页
宋喜梅%叶冰莹%林荣华%代容春%何文锦%陈由强%陈如凯
宋喜梅%葉冰瑩%林榮華%代容春%何文錦%陳由彊%陳如凱
송희매%협빙형%림영화%대용춘%하문금%진유강%진여개
蔗糖磷酸合成酶基因%定点突变%中间缺失突变体%模序
蔗糖燐痠閤成酶基因%定點突變%中間缺失突變體%模序
자당린산합성매기인%정점돌변%중간결실돌변체%모서
sucrose phosphate synthase gene%middle fragment deletion mutant%site-directed mutagenesis%motifs
根据NCBI中EU278617序列设计引物,利用甘蔗SPSⅢG2菌液为模板,扩增SPSⅢ8-11并克隆到T载体后测序.通过排除野生型DNA的定点突变PCR方法获得SPSⅢ8-11中第8内含子缺失TGCATG的突变体pMD-SPSⅢ8-11-A,酶切后测序鉴定.成功构建了甘蔗SPSⅢ8-11中间缺失突变体pMD-SPSⅢ8-11-A.
根據NCBI中EU278617序列設計引物,利用甘蔗SPSⅢG2菌液為模闆,擴增SPSⅢ8-11併剋隆到T載體後測序.通過排除野生型DNA的定點突變PCR方法穫得SPSⅢ8-11中第8內含子缺失TGCATG的突變體pMD-SPSⅢ8-11-A,酶切後測序鑒定.成功構建瞭甘蔗SPSⅢ8-11中間缺失突變體pMD-SPSⅢ8-11-A.
근거NCBI중EU278617서렬설계인물,이용감자SPSⅢG2균액위모판,확증SPSⅢ8-11병극륭도T재체후측서.통과배제야생형DNA적정점돌변PCR방법획득SPSⅢ8-11중제8내함자결실TGCATG적돌변체pMD-SPSⅢ8-11-A,매절후측서감정.성공구건료감자SPSⅢ8-11중간결실돌변체pMD-SPSⅢ8-11-A.
Designed primers according to EU278617 of NCBI. Amplified the target SPSⅢ8-11 by PCR protocol from sugarcane SPSⅢG2, then cloned into pMD18-T vector and sequenced. The middle fragment deleted mutant of SPSⅢ8-11 (pMD-SPSⅢ8-11-A) was induced by PCR-based site-directed mutagenesis, digested and sequenced.Middle fragment deleted mutant of SPSⅢ8-11 from sugarcane (pMD-SPSⅢ8-11-A) was constructed successfully.
