|
|
|
| Gene Knocking Out of Hyaluronidase in Streptococcus zoopidemics |
| CUI Ya-na,SU Xu-dong,WANG Yu,MA Xiao-yan,WANG Xue-jing,LIN Yang,ZHANG Wei |
| College of Food Science and Technology,Agricultural University of Hebei,Baoding 071001,China |
|
|
|
Abstract To construct knockout vectors containing ampicillin resistant gene and partial sequence of hyaluronidase gene(Hyl)so that Hyl can be knock out by transforming the plasmid into Streptococcus zoopidemics mutans.First,partial sequence of Hyl（Hyl-1）was cloned into the vector of pMD19-T by using DNA of Streptococcus zoopidemics as template,and then a knockout vector pMD19T-SA was constructed, in which Hyl-1 gene was disrupted by inserting ampicillin resistant gene(Amp)from reverse PCR. As expected, the vector was proved to be consisted of Hyl-1-Amp-Hyl-1-pMD19-T.Thereafter,DNA fragment of Hyl-1-Amp-Hyl-1 was subcloned into pBR322 vector,the resulting construct was then checked by PCR and restriction analysis for the proper configuration of the knockout vector pBR322-SA.Both of the knockout vectors were used to transform Streptococcus zoopidemics and one recombinant was obtained in result. From results of PCR and Hyl activity assay, it was indicated that in the recombinant the Hyl gene was disrupted completely.
|
|
Received: 09 September 2009
Published: 21 December 2009
|
|
|
|
|
| ［1］ |
朱广华,方煜平.透明质酸制备方法及应用研究的进展.中国医药工业杂志, 1996, 2(8):382～384 Zhu G H, Fang Y P.Chinese Journal of Pharmaceuticals, 1996, 2(8):382～384
|
|
|
| ［2］ |
Takashi Yamada,Takeru Kawasaki. Microbial synthesis of hyaluronan and chitin. New Approaches, 2005,99(6):521～528
|
|
|
| ［3］ |
Farrell A M,Taylor D, Holland K T. Cloning nucleotide sequence determination and expression of the Staphylococcus aureus hyaluronate lyase gene. FEMS Microbiol Lett,1995, 130(1):81～85
|
|
|
| ［4］ |
Berry A M, Lock R A,Thomas S M, et al. Cloning and nucleotide sequence of the Streptococcus pneumoniae hyaluronidase gene and purification of the enzyme from recombinant Escherichia coli. Infect Immun,1994, 62(3):1101～1108
|
|
|
| ［5］ |
谢承佳,何冰芳,李霜. 基因敲除技术及其在微生物代谢工程方面的应用.生物加工过程杂志，2007，5（3）：10～14 Xie C J, He B F, Li S.Chinese Journal of Bioprocess Engineering, 2007，5（3）:10～14
|
|
|
| ［6］ |
田琳，李玉，史文玉，等. 分枝杆菌甾酮C1，2位脱氢酶基因敲除的研究。中国生物工程杂志，2007，27（5）：39～44 Tian L, Li Y, Shi W Y, et al.China Biotechnology, 2007, 27(5): 39~44
|
|
|
| ［7］ |
饶志明，张君胜，沈 微，等. 谷氨酸棒杆菌高丝氨酸脱氢酶编码基因hom的敲除。中国生物工程杂志, 2006, 27(1)：59～63 Rao Z M, Zhang J S, Shen W, et al.China Biotechnology, 2006, 27(1): 59～63
|
|
|
| ［8］ |
Ju A P, Wang C J, Li M, et al. Construction of RevS gene knockout mutant of Streptococcus suis serotype 2.Zhonghua Liu Xing Bing Xue Za Zhi,2008, 29(1): 59～64
|
|
|
| ［9］ |
Chien L J, Lee C K. Hyaluronic acid production by recombinant Lactococcus lactis. Microbiol Biotechnol, 2007,77(2):339～346
|
|
|
| ［10］ |
张云开.透明质酸生产菌代谢工程研究. 南宁：广西大学，2002 Zhang Y K. Metabolic Engineering Research of Hyaluronic AcidProducing Bacterium Streptococcus equi.Nanning Guangxi University, 2002
|
|
|
| ［11］ |
郝宁，张晋宇，陈国强. 在兽疫链球菌中表达vgb基因好HA合成基因提高透明质酸产量.中国生物工程杂志，2005，25（6）：56～60 Hao N, Zhang J Y, Chen G Q.China Biotechnology，2005，25（6）：56～60
|
|
|
| ［12］ |
黄艳. 一株透明质酸生产菌的鉴定及其透明质酸分解酶基因的克隆和鉴定. 南宁： 广西大学, 2004 Huang Y. Hyaluronic Acid and the Clonging and Expression of its Hyalurondiase Gene.Nanning Guangxi University, 2004
|
|
|
| ［13］ |
石艳丽，郭学平，栾贻宏. 产透明质酸菌的育种概况.食品与药品，2006，8(10)22～24 Shi Y L, Guo X P, Luan Y H.Food and Drug, 2006，8(10)22～24
|
|
|
| ［14］ |
Daisuke Takamatsu,Makoto Osaki,Tsutomu Sekizaki. Construction and characterization of Streptococcus suisEscherichia coli shuttle cloning vectors. Plasmid, 2001,(45):101～113
|
|
|
| ［15］ |
Daisuke Takamatsu,Makoto Osaki,Tsutomu Sekizaki.Thermosensitive suicide vectors for gene replacement in Streptococcus suis. Plasmid,2001,(46):140～148
|
|
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
