摘要
在提取真菌互格链格孢(Alternaria alternata)总RNA的基础上,通过一步法RT-PCR技术,获得了多聚半乳糖醛酸酶(PGase)的cDNA,并将其克隆至载体pBluescript SK(-),经双酶切和测序分析后,通过Gen-Bank对其进行序列比对分析。结果表明该cDNA序列与已公布的序列存在14个碱基的差异,密码子简并性只导致3个氨基酸的不同,但不影响PGase与多聚半乳糖醛酸酶抑制蛋白(PGIP)的结合能力。研究结果为利用酵母双杂交技术来获得高活性的植物PGIP打下了基础。
The cDNA of polygalacturonase （PGase） from fungus Alternaria alternata was cloned by RT-PCR after extracting its total RNA, then was ligated into vector pBluescript SK （ - ）. After double-digestion identification and sequencing analysis, the cDNA was aligned with published sequences via GenBank, which reveals that there is a difference of fourteen bases and three amino acids for the codon degeneracy, however, this will not affect the ability of the PGase to bind with PGIP. The result will establish a foundation for obtaining high activity PGIP by the method of yeast two-hybrid.
出处
《东北林业大学学报》
CAS
CSCD
北大核心
2006年第6期84-86,113,共4页
Journal of Northeast Forestry University
基金
中南林业科技大学青年科学基金重点项目(05007A)
湖南省教育厅科学研究项目(05C325)。
