Cloning and Characterization of a Copper/Zinc-Superoxide Dismutase Gene from Aspergillus Japonicus

麴菌銅鋅型超氧歧化酶基因選殖

林棋財(Chi-Tsai Lin) ； 林銘澤(Ming-Tse Lin) ； 許垤棊(Dey-Chyi Sheu) ； 段國仁(Kow-Jen Duan)

《TAIWANIA》 39卷1＆2期 (1994/10) Pp. 73-79

https://doi.org/10.6165/tai.1994.39.73

摘要

依據玉米Cu/Zn-SOD之核酸序列，合成兩股引子，並以麴菌cDNA當模版，以PCR方法增生出一段0.3kb DNA，經次選殖及定序，其序列與玉米Cu/Zn-SOD之序列同質性高達73%，進一步以此0.3kb DNA當探針，進行麴菌基因庫之篩選，結果篩得一株含有0.4kb之正反應株，經定序分析，發現可推導出含120個胺基酸序列，其序列和其他生物Cu/Zn-SOD之序列同質性很高，並含有與銅和鋅進行相對應（coordinating）之胺基酸序列。

並列摘要

In order to clone a Cu/Zn-SOD gene from Aspergillus japonicus, the highly conserved regions of the SOD sequence of maize and other species were compared and two oligonucleotides were synthesized. The Aspergillus japonicus cDNA was then used as the template to get a 0.3 kb fragment by PCR technique. The 0.3kb DNA fragment has high homology compared with maize SOD-4 cDNA. Then we used the 0.3kb fragment as probe to screen Aspergillus japonicus genomic libraries. A 0.4 kb DNA frgment from purified positive plaque was subcloned into pGEM-7zf(+) vector and sequenced. Nucleotide sequence analysis of this clone revealed that it comprised an open reading frame coding for 120 amino acid residues. The encoded polypeptide is highly homologous with other plant SODs. The residues required for coordinating copper and zinc are also conserved as they are among all reported Cu/Zn-SOD sequences.

並列關鍵字

Aspergillus japonicus ； copper/zinc-superoxide dismutase (Cu/Zn-SOD)