Cloning and expression of a chicken alpha-amylase gene

被引：15

作者：

Benkel, BF ^{[1
]}

Nguyen, T ^{[1
]}

Ahluwalia, N ^{[1
]}

Benkel, KI ^{[1
]}

Hickey, DA ^{[1
]}

机构：

[1] UNIV OTTAWA,DEPT BIOL,OTTAWA,ON K1N 6N5,CANADA

来源：

GENE | 1997年 / 192卷 / 02期

基金：

加拿大自然科学与工程研究理事会;

关键词：

nucleotide sequence; gene structure; transgenic yeast;

D O I：

10.1016/S0378-1119(97)00102-9

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We have isolated and sequenced a genomic clone for a pancreatic a-amylase gene (amy) of the chicken (Gallus gallus). The gene is interrupted by nine introns, spans over 4 kb, and encodes a protein (AMY) of 512 aa that is 83% identical to the human pancreatic alpha-amylase enzyme. Southern blot analysis of chicken DNA revealed two distinct pancreatic amy loci. In addition, we have generated a cDNA from chicken pancreatic RNA corresponding to the coding sequence of the genomic clone. The cDNA was inserted into a yeast expression vector, and the resulting construct used to transform Saccharomyces cerevisiae cells. Transformed yeast cells synthesized and secreted active AMY enzyme, and the gel migration pattern of the cl-amylase produced by the yeast cells was identical to that of the native chicken enzyme. (C) 1997 Elsevier Science B.V.

引用

页码：261 / 270

页数：10

共 33 条

[1] PCR AMPLIFICATION OF UP TO 35-KB DNA WITH HIGH-FIDELITY AND HIGH-YIELD FROM LAMBDA-BACTERIOPHAGE TEMPLATES [J].