摘要
为进一步探讨苯丙氨酸解氨酶(PAL)基因的表达和研究其功能,采用通用植物总RNA提取试剂盒提取蓝莓果实总RNA,并根据同源基因设计相关引物,利用反转录PCR方法克隆苯丙氨酸解氨酶PAL编码基因,将其连接到pMD18-T载体上构建克隆载体并酶切鉴定。然后将PAL目的基因连接到pET-20b(+)上构建表达载体,并通过PCR检测和酶切双重方法鉴定。结果表明,成功获得了258bp大小的PAL片段,并成功构建表达载体。
To deeply explore the expression and function of gene PAL,total RNA of blueberry fruit was extracted 6 by extracting kit in universal plant total RNA,and related primers were designed according to the homologous genes,coding gene of phenylalanine ammonialyase was cloned using reverse transcription PCR,and then was connected to pMD18-T vector to construct cloning vector and identify by restriction enzyme analysis method.Then PAL gene was ligated to pET-20b（＋）vector to acquire expression vector,and both methods of restriction enzyme analysis and PCR was conducted for detection.The result indicated that 258 bp band of PAL was got and the expression vector was established successfully.
出处
《贵州农业科学》
CAS
2015年第12期26-29,共4页
Guizhou Agricultural Sciences
基金
吉林农业科技学院专项基金项目"长白山重点实验室"[吉农院合字(2012)第714号]
关键词
蓝莓
苯丙氨酸解氨酶
克隆
表达载体
blueberry
phenylalanine ammonialyase
clone
expression vector
