In this paper,the cloning of a β-xylanase structural gene and its 5′flanking regions gene from Aspergillus niger was studied.On the base of nucleic acid DNA sequence of β-xylanase gene,a pair of degenerate primers was designed.Using the genomic DNA of Aspergillus niger as template,a 1.6 kb fragment was amplified by PCR and cloned into the vector pBS-T.Sequence analysis showed that the structural gene of β-xylanase is 744 bp length including a 66 bp intron and a 678 bp encoding region,which encoded a polypeptide of 225 amino acids and included the signal sequence of 18 amino acids.The homology analysis of the xynB cDNA sequence shares 98% in base sequence with the β-xylanase gene in GeneBank and the amino acid sequence shares 99%.Upstream of the tranlation start site,there were classic core element of Eukaryotic Gene Promoter,such as transcription start site,TATA box and CAAT box elements.The work was beneficial in construction of homologus expression for xylanase efficient expression vector.
BAI Ai-zhi
,
YAN Zu-wei
,
TANG Guo-min
,
LIANG Yun-zhang
. Cloning and Sequence Analysis of β-xylanase Structural Gene and the 5′Flanking Regions Gene from Aspergillus niger[J]. Acta Agriculturae Boreali-Sinica, 2009
, 24(5)
: 73
-76
.
DOI: 10.7668/hbnxb.2009.05.016
[1] 张红莲,姚 斌,范云六.木聚糖酶的分子生物学及其应用[J].生物技术通报,2002(3):23-26.
[2] Sunna A,Antramikian C.Xylanolytic enzymes from fungi and bacteria[J].Critical Rev Biotechnol,1997,17:39-67.
[3] Beg Q K,Kapoor M,Mahajan L,et al.Microbial xylanases and their industrial applications:a review[J].Appl Microbiol Biotechnol,2001,56:326-338.
[4] 毛连山,勇 强,宋向阳,等.内切木聚糖酶的选择性纯化及酶解制备低聚木糖的研究[J].林产化学与工业,2006,26(1):124-126.
[5] Subramaniyan S,Prema P.Biotechnology of microbial xylanases:enzymology,molecular biology,and application[J].Crit Rev Biotechnol,2002,22(1):33-64.
[6] Khandeparkar R,Bhosle N B.Application of thermoalkalophilic xylanase from Arthrobacter sp.MTCC 5214 in biobleaching of kraft pulp[J].Bioresource Technology,2007,98:897-903.
[7] Vazquez M J,Alonso J L,Dominguez H,et al.Enhancing the potential of oligosaccharides from corncob autohydrolysis as prebiotic food ingredients[J].Industrial Crops and Products,2006,24:152-159.
[8] Eun J S,Beauchemin K A,Hong S H,et al.Exogenous enzymes added to untreated or ammoniated rice straw:Effects on in vitro fermentation characteristics and degradability[J].Animal Feed Science and Technology,2006,131:86-101.
[9] Ohgren K,Bura R,Saddler J,et al.Effect of hemicellulose and lignin removal on enzymatic hydrolysis of steam pretreated corn stover[J].Bioresource Technology,2007,98:2503-2510.
[10] 聂国兴,王俊丽,明 红.木聚糖酶的应用现状与研发热点[J].工业微生物,2008,38(1):53-59.
[11] 张世敏,刘 寅,刘新育,等.木聚糖酶基因研究进展[J].微生物学杂志,2006,26(4):61-67.
[12] 朱 衡,瞿 峰,朱立煌.利用氯化苄提取适于分子生物学分析的真菌DNA[J].真菌学报,1994,13(1):34-40.
[13] 萨姆布鲁克.分子克隆实验指南[M].第2版.北京:科学出版社,1992.
[14] Nagata O.The Aspergillus nidulans nuclear proteins bind to a CCAAT element and the adjacent upstream sequence in the promoter region of the starch-inducible Taka-amylase A gene[J].Mol Genet,1993,237:251-260.
[15] 朱兴国,Wang H M,仇润祥,等.黑曲霉糖化酶cis调控区上两个CCAAT框协同参与基因启动子的转录激活作用[J].中国科学(C辑),2003,33(6):495-504.
