In our experiment , the specific fragment was amplified from transgenic bobwhite genome dna at annealing temperature 61 by using high - fidelity pfu dna polymerase and cloned into clone vector pgem - 7fz ( + ) , then sequenced . the cloned sequence was completely identical to the sequence which was issued in genbank 本实验采用了高保真pfudna聚合酶,在退火温度61条件下从转基因bobwhite品种基因组dna中扩增出特异性片段,将此片段插入克隆载体pgem - 7fz ( + ) ,经测序和序列分析表明,所扩增得到的片段含有bar基因完整的读码框,并且序列与genbank中发表的序列完全一致。
2.
In the experiment , the full code sequence of bar gene was cloned by pcr from transgenic herbicide resistant bobwhite wheat and checked . it was expressed in e . coli and its protein was determined . after having been properly modified , the bar gene which correctly codes pat was cloned into binary vector pbi121 and then transferred into lba4404 by triparantal crossing , which is the prerequisite work for genetic transformation 本实验从抗除草剂转基因bobwhite小麦中,利用pcr克隆的方法扩增出bar基因全长,并在原核表达系统中表达,鉴定表达蛋白的活性,将能够正确编码ppt乙酰转移酶的bar基因片段,经过适当的修饰构建入真核表达载体。