[Construction of a set of secreting expression vectors for Saccharomyces cerevisiea].
The DNA fragment ecoding the Signal peptide of inulinase of Kluyveromyces smarxianu was synthesized chemically. This fragment was cloned in-frame in the expression vector pYES2 of Saccharomyces cerevisiae, resulting in a set of new secreting expression vectors pYES2 I, pYES2 II, pYES2 III. The L-Asparaginase gene (ASN) of E. coli and alpha-acetylactate decarboxylase gene (ALDC) of B. brevis which were amplified by PCR and cloned into the new vectors respectively were transformed into Saccharomyces cerevisia, and most of enzyme activities were secreted into the medium. The new secreting expression vectors still have excellent segregational stability even after growth for 100 h in the absence of selective pressure.
Zhao
Yingyi
Y
Institute of Food and Biology Engineering, South China Science and Technology University, Guangzhou 510640, China.
Liang
Shizhong
S
Huang
Kun
K
Huang
Ribo
R
D001215:Asparaginase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001407:Bacillus / Q000737:chemistry; D002262:Carboxy-Lyases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000737:chemistry; D015870:Gene Expression; D005822:Genetic Vectors; D006026:Glycoside Hydrolases / Q000235:genetics / Q000302:isolation & purification; D007716:Kluyveromyces / Q000737:chemistry; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; C023086:inulinase; D001215:Asparaginase; D002262:Carboxy-Lyases; C023746:acetolactate decarboxylase