孝感学院学报
孝感學院學報
효감학원학보
JOURNAL OF XIAOGAN UNIVERSITY
2009年
3期
10~14
,共null页
李长春 王志华 乔富兴 熊燃
李長春 王誌華 喬富興 熊燃
리장춘 왕지화 교부흥 웅연
海甘蓝 硫甙 Thiohydroximate S-glucosyltransferase 外显子拼接 表达载体构建
海甘藍 硫甙 Thiohydroximate S-glucosyltransferase 外顯子拼接 錶達載體構建
해감람 류대 Thiohydroximate S-glucosyltransferase 외현자병접 표체재체구건
Crambe abyssinica ; glucosinolate; thiohydroximate S-glucosyltransferase; extron assembly; expression vector construc
Thiohydroximate S-glucosyltransferase（S-GT）是硫甙生物合成的一个关键酶。根据发表的甘蓝型油菜S—GT基因cDNA序列设计引物，以海甘蓝总DNA为模板进行PCR扩增，获得S-GT基因全长，构建了海甘蓝S-GT基因的植物双元表达载体pCambia2300sn-CaSGT。用PCR的方法对CaSGT基因的外显子进行了拼接，将得到的序列正向插入到了大肠杆菌表达载体pET-32b（＋）中，得到重组质粒pET-32b（＋）-CsS—GT，为进一步研究CaSGT的生物学特性打下基础。
Thiohydroximate S-glucosyltransferase（S-GT）是硫甙生物閤成的一箇關鍵酶。根據髮錶的甘藍型油菜S—GT基因cDNA序列設計引物，以海甘藍總DNA為模闆進行PCR擴增，穫得S-GT基因全長，構建瞭海甘藍S-GT基因的植物雙元錶達載體pCambia2300sn-CaSGT。用PCR的方法對CaSGT基因的外顯子進行瞭拼接，將得到的序列正嚮插入到瞭大腸桿菌錶達載體pET-32b（＋）中，得到重組質粒pET-32b（＋）-CsS—GT，為進一步研究CaSGT的生物學特性打下基礎。
Thiohydroximate S-glucosyltransferase（S-GT）시류대생물합성적일개관건매。근거발표적감람형유채S—GT기인cDNA서렬설계인물，이해감람총DNA위모판진행PCR확증，획득S-GT기인전장，구건료해감람S-GT기인적식물쌍원표체재체pCambia2300sn-CaSGT。용PCR적방법대CaSGT기인적외현자진행료병접，장득도적서렬정향삽입도료대장간균표체재체pET-32b（＋）중，득도중조질립pET-32b（＋）-CsS—GT，위진일보연구CaSGT적생물학특성타하기출。
Thiohydroximate S-glucosyltransferase （S-GT） plays a key role in the process of glucosinolate （GS） biosynthesis. Full-length CaGST gene was amplified from Crarnbe abyssinica genomic DNA according to the cDNA sequence of S-GT gene in Brassica napus, and then the sequence was cloned into plasmid pCambia2300sn and located between 35S promoter and NOS terminater, Meanwhile, we assembled the two extrons of CaSGTgene by PCR strategy and cloned the sequence into pET-32b （＋） to get the recombinant expression vector PET-32b （＋）-CaSGT. tion
