摘要
采用Ｌａｍｂｄａｇｔ１０载体构建了盐藻ｃＤＮＡ文库，文库大小为１０６／ｍＬ，插入片段平均长度大于１ｋｂ．根据果蝇、兔、鼠编码其３－磷酸甘油脱氢酶基因的辅酶ＮＡＤ的结合功能区保守序列设计一对引物，大小分别为２１ｂｐ和１８ｂｐ．ＰＣＲ扩增得到与果蝇相同的２９０ｂｐ特异扩增片段，以该片段为探针，采用缺口平移系统标记，原位杂交。
The cDNA library (106 clones) of Dunaliella Salina was obtained using a lambda gt10 vector.The average size of insertion is more than 1kb.Comparing the conservative sequence encoding NAD binding region of glycerol3phosphate dehydrogenase among Drosophila,rabbit and mouse,we designed and synthesized a pair of primers.The size of the primers were 21bp and 18bp respectively.PCR amplification was performed and obtained ene specific amplified fragment of 290bp which is as same as in Drosophila DNA.The fragment was recoveried and used as the probe for screening glycerol3phosphate dehyogenase gene from cDNA library.Nick translation system was used for labelling probe.As a result of in situ hybrdization,36 positive clones were obtained.
出处
《四川大学学报（自然科学版）》
CAS
CSCD
北大核心
1997年第6期839-842,共4页
Journal of Sichuan University(Natural Science Edition)
基金
国家自然科学基金
关键词
盐藻
磷酸甘油脱氢酶
CDNA文库
绿藻
基因
Dunaliella Salina,glycerol3phosphate dehydrogenase,cDNA library,positive colne
