修饰合成冬鲽抗冻肽基因的设计和克隆  

Design and Cloning a Modified Synthetic Gene for Flounder Antifreeze Peptide

作　　者：达来[1] 朱晔荣[1] 马振毅[1] 王非[1] 阚锐[1] 

机构地区：[1]内蒙古大学生物学系,内蒙古呼和浩特010021

出　　处：《内蒙古大学学报（自然科学版）》2000年第2期216-222,共7页Journal of Inner Mongolia University：Natural Science Edition

基　　金：国家自然科学基金!(39660045)

摘　　要：:动物界和植物界密码子使用频率不同 .冬鲽抗冻肽中丙氨酸占 60 %而且偏爱GCC ,3 8个Ala密码子中 2 3个为GCC .我们设计合成抗冻肽基因时采用了植物基因常用密码子 .用甘薯(Ipomoeabatatas)信号肽序列取代了抗冻肽的pre序列 ,省略了Pro序列 ,并构建了信号肽下游二、四、八拷贝正向串联的成熟肽的修饰基因 ,经测序证实与设计完全一致 .利用QIA表达系统 ,在大肠杆菌中实现了DHFRCodon usage varies throughout the animal and plant kingdoms.The flounder antifreeze peptide (AFP) is Ala rich (60%) and the Ala codons of the fish cDNA reveal a strong bias toward GCC (23/38).We have taken the opportunity to modify the codon selection for all the amino acids to be consistent with normal usage in plant genes.The pre sequence of AFP was replaced by the pre sequence of sweet potato Sporamin A (EMBL X13 509),and the pro sequence was not included.The modified genes coding 2,4,8 tandem mature AFP were constructed and confirmed by DNA sequencing.Using QIA express system,expression of DHFR mature AFP constructs in E.coli suggests the modified synthetic AFP gene could be work in plants.

关 键 词：抗冻肽 合成基因 冬Die 修饰基因 设计 克隆 

分 类 号：Q78[生物学—分子生物学] Q514.3