Biopolym. Cell. 1995; 11(2):39-45.
Структурно-функціональний аналіз промоторної області гена алкогольоксидази метилотрофних дріжджів Hansenula polymorpha
1Витвицька О. П., 1Злочевський М. Л., 1Бебуров М. Ю.
  1. Відділення регуляторних систем клітини Інституту біохімії ім. О. В. Палладіна НАН України
    вул. Драгоманова 14/16, Львів, Україна, 79005


Досліджено вплив делецій в промоторі гена АОХ на експресію генів lac Z Escherichia coli і АОХ Н. polymorpha. Показано, що область між –403 п. о. і кодоном ATG про­ мотора АОХ абсолютно необхідна для транскрипції гена і її регуляції у метилотрофних дріжджах. Встановлено присутність у промоторі ділянок, активуючих експресію. Виявлено відмінності в регуляції експресії генів, що знаходяться під контролем промотора АОХ, у дріжджах Н. polymorpha і Saccharomyces cerevisiae.


[1] Cregg JM, Vedvick TS, Raschke WC. Recent advances in the expression of foreign genes in Pichia pastoris. Biotechnology (N Y). 1993;11(8):905-10.
[2] Janowicz ZA, Melber K, Merckelbach A, Jacobs E, Harford N, Comberbach M, Hollenberg CP. Simultaneous expression of the S and L surface antigens of hepatitis B, and formation of mixed particles in the methylotrophic yeast, Hansenula polymorpha. Yeast. 1991;7(5):431-43.
[3] Veale RA, Giuseppin ML, van Eijk HM, Sudbery PE, Verrips CT. Development of a strain of Hansenula polymorpha for the efficient expression of guar alpha-galactosidase. Yeast. 1992;8(5):361-72. Erratum in: Yeast 1992 Sep;8(9):813.
[4] Veenhuis M, Van Dijken JP, Harder W. The significance of peroxisomes in the metabolism of one-carbon compounds in yeasts. Adv Microb Physiol. 1983;24:1-82.
[5] Roggenkamp R, Janowicz Z, Stanikowski B, Hollenberg CP. Biosynthesis and regulation of the peroxisomal methanol oxidase from the methylotrophic yeast Hansenula polymorpha. Mol Gen Genet. 1984;194(3):489-93.
[6] Pfeifer K, Prezant T, Guarente L. Yeast HAP1 activator binds to two upstream activation sites of different sequence. Cell. 1987;49(1):19-27.
[7] West RW Jr, Yocum RR, Ptashne M. Saccharomyces cerevisiae GAL1-GAL10 divergent promoter region: location and function of the upstream activating sequence UASG. Mol Cell Biol. 1984;4(11):2467-78.
[8] Beier DR, Sledziewski A, Young ET. Deletion analysis identifies a region, upstream of the ADH2 gene of Saccharomyces cerevisiae, which is required for ADR1-mediated derepression. Mol Cell Biol. 1985;5(7):1743-9.
[9] Bram RJ, Kornberg RD. Specific protein binding to far upstream activating sequences in polymerase II promoters. Proc Natl Acad Sci U S A. 1985;82(1):43-7.
[10] Guarente L. UASs and enhancers: common mechanism of transcriptional activation in yeast and mammals. Cell. 1988;52(3):303-5.
[11] Nehlin JO, Carlberg M, Ronne H. Control of yeast GAL genes by MIG1 repressor: a transcriptional cascade in the glucose response. EMBO J. 1991;10(11):3373-7.
[12] Ledeboer AM, Edens L, Maat J, Visser C, Bos JW, Verrips CT, Janowicz Z, Eckart M, Roggenkamp R, Hollenberg CP. Molecular cloning and characterization of a gene coding for methanol oxidase in Hansenula polymorpha. Nucleic Acids Res. 1985;13(9):3063-82.
[13] Godecke S, Hollenberg CP. In vitro and in vivo DNA: protein interactions at the MOX-promoter of the methylotrophic yeast Hansenula polymorpha 11 Abstr. of 15th Int conf yeast genet mol biol. Banff, 1990: 280.
[14] Maniatis T, Fritsch EF, Sambrook J. Molecular cloning: a laboratory manual. New York: Cold Spring Harbor Lab, 1982; 545 p.
[15] Ish-Horowicz D, Burke JF. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981;9(13):2989-98.
[16] Sherman F, Fink G, Hicks J. Methods in Yeast Genetics: A Laboratory Course Manual. New York : Cold Spring. Harbor Lab., 1987. 180 p.
[17] Sambrook J, Fritsch EE, Maniatis T. Molecular cloning. Cold Spring Harbor Lab. press, 1989; 625 p.
[18] Ito H, Fukuda Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983;153(1):163-8.
[19] Rose M, Botstein D. Construction and use of gene fusions to lacZ (beta-galactosidase) that are expressed in yeast. Methods Enzymol. 1983;101:167-80.
[20] Sibirny? AA, Titorenko VI. A method of quantitative determination of alcohol oxidase and catalase in yeast colonies. Ukr Biokhim Zh. 1986;58(5):65-8.
[21] Miller JL. Experiments in molecular genetics. New York: Cold Spring Harbor Lab., 1972; 468 p.
[22] Sibirny AA, Titorenko VI, Efremov BD, Tolstorukov II. Multiplicity of mechanisms of carbon catabolite repression involved in the synthesis of alcohol oxidase in the methylotrophic yeastPichia pinus. Yeast. 1987;3(4):233–41.
[23] Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193(1):265-75.
[24] Myers AM, Tzagoloff A, Kinney DM, Lusty CJ. Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene. 1986;45(3):299-310.
[25] Gracheva VV, Mikhaylover VM, Shchedrin AV. et al. Preconditions for creation gene expression system in the methylotrophic yeast Hansenula polymorpha. Cloning of methanol oxidase. Biotechnology, medicine and the national economy: Coll. scientific. paper. VNIISENTI. M.; 1990. 5-12.
[26] Didion T, Roggenkamp R. Deficiency of peroxisome assembly in a mutant of the methylotrophic yeast Hansenula polymorpha. Curr Genet. 1990;17(2):113–7.
[27] Buckholz R, Lin YC, Koutz P. Upstream sequences required for methanol. glucose regulation of the P. pastoris alcohol oxidase gene. Abstr. of 13th Int conf yeast gen mol biol. Banff. 1986. Vol. 2: 45.