酵母菌杂交

一、实验原理
酵母菌的生活史属于双单性类型，它们的单倍体细胞和二倍体细胞一般都能进行无限制的分裂。以单倍体细胞为主的酵母菌称为单倍体酵母，例如粟酒裂殖酵母（Schizosaccharamycespombe）。以二倍体细胞为主的酵母菌称为二倍体酵母，如啤酒酵母（又称面包酵母，Saccharomycescerevisiae）。这两种酵母常用作遗传学研究的材料，而啤酒酵母在酿酒工业和食品工业中又有很大的应用价值。本实验用异宗配合型的啤酒酵母的单倍体菌株进行杂交实验。任何一株酵母菌，如果使二倍体的营养细胞处于形成孢子的条件下，都会发生减数分裂，生成四分体和子囊。每个子囊孢子具有四分子核，正常情况下子囊中有4个子囊孢子，其中有2个a和2个α，a和α表示两种相对的接合型。具有这两种接合型的子囊孢子萌发生长成单倍体的菌株，不同接合型的单倍体菌株细胞接合生成二倍体菌株。具有这样生活史的酵母菌称为异宗配合型酵母。异宗配合型啤酒醇母的生活史见图11－1。
单倍体菌株分属a和α两种接合型，是受一对等位基因a和α控制的，这对等位基因的座位在第三连锁群靠近着丝粒的地方。在光学显微镜下观察酵母的染色体是十分困难的，但借助于电子显微镜，对酵母菌的细胞学已了解得较多了。现已知啤酒酵母具有17个连锁群（n=17）。
两个属于不同接合型的营养缺陷型的酵母单倍体菌株，在基本培养基上都不能形成菌落。当它们杂交形成二倍体杂种细胞，就能在基本培养基上生长。二倍体杂种细胞在形成子囊孢子过程中，发生染色体重组，而使产生的子囊孢子出现重组性状。
二、实验目的
通过酵母菌杂交实验了解异宗配合型啤酒酵母的生活史和基因自由组合定律，掌握酵母菌杂交的基本方法。
三、实验材料
1.啤酒酵母单倍体腺嘌呤缺陷型（ade-），接合型为a；
2.啤酒酵母单倍体组氨酸缺陷型（Hisl-），接合型为α。
四、实验器具和药品
1.用具：试管，离心管，离心机，吸管，培养皿，三角烧瓶，涂布棒，石英砂。
2.药品：生理盐水（0.85克NaCl溶于100毫升蒸馏水中，15磅压力下消毒15分钟），纤维素酶或蜗牛酶。
3.培养基：
（1）完全液体培养基：蛋白胨2克，酵母浸母汁1克，葡萄糖2克，水100毫升，pH6.0，8磅压力下灭菌30分钟。
（2）完全固体培养基：在完全液体培养基中加2％琼脂。
（3）基本液体培养基：葡萄糖10克，（NH4）2SO41克，K2HPO40.125克，KH2PO40.875克，KI*母液1毫升，MgSO4·7H2O0.5克，CaCl2·2H2O0.1克，NaCl0.1克，微量元素母液**1毫升，维生素母液***1毫升，水1000毫升，pH5.3，8磅压力下灭菌30分钟。
*KI母液：10毫克KI于100毫升水中。
**微量元素母液：H3PO41毫克，ZnSO4·7H2O7毫克，CuSO4·5H2O1毫克，CoCl2·6H2O5毫克，水100毫升。
***维生素母液：烟碱酸40毫克，维生素B140毫克，肌醇200毫克，核黄素20毫克，对-氨基苯甲酸20毫克，吡哆醇40毫克，泛酸40毫克，生物素0.2毫克，水100毫升。
（4）基本固体培养基：在基本液体培养基中加2％琼脂。
（5）产孢子培养基：CH3COONa8.2克，KCl1.86克，吡哆酵母液*1毫升，泛酸母液**1毫升，生物素母液***1毫升，琼脂20克，蒸馏水1000毫升。

德国默克全系列 现货促销产品

BPHHSP-BASEWidescreen? BeadPlex? Human Heat Shock Protein Panel -Base
BPHHSP-H27Human Heat Shock Protein 27
BPHHSP-H60Heat Shock Protein 60 (HSPD1)
BPHHSP-H70Heat Shock Protein 70 (HSP72, HSPA1A&A1B
BPHHSP-H90Heat Shock Protein 90 alpha (HSPAA1)
BPHHSP-P27Phospho-Heat Shock Protein 27 (Ser78/Ser82)
03-32-0009-100MGELASTASE SUBSTRATE VIII, COLORIMETRIC
03-32-1501-25MGZ-PHE-ARG-7-AMIDO-4-METHYLCOUMARIN, HCL
03-32-1501-5MGZ-PHE-ARG-7-AMIDO-4-METHYLCOUMARIN, HCL
03-32-5032-1MGMMP-2/MMP-7 SUBSTRATE, FLUOROGENIC
03-32-5033-1MGMMP-2/MMP-7 SUBSTRATE CONTROL
03-34-0001-25MGH-GLY-PRO-ARG-PRO-OH
03-34-0001-5MGH-GLY-PRO-ARG-PRO-OH
03-34-0002-25MGH-ARG-GLY-ASP-SER-OH
03-34-0027-25MGH-GLY-ARG-GLY-ASP-SER-OH
03-34-0035-25MGH-GLY-ARG-GLY-ASP-SER-PRO-OH
03-34-0035-5MGH-GLY-ARG-GLY-ASP-SER-PRO-OH
03-34-0051-100MGCALPEPTIN
03-34-0051-25MGCALPEPTIN
03-34-0051-5MGCALPEPTIN
03-34-0052-25MGH-GLY-ARG-ALA-ASP-SER-PRO-OH
03-34-0052-5MGH-GLY-ARG-ALA-ASP-SER-PRO-OH
03-34-0055-25MGH-GLY-ARG-GLY-ASP-THR-PRO-OH
03-34-0055-5MGH-GLY-ARG-GLY-ASP-THR-PRO-OH
05-22-0930-100GMB-ENDORPHIN (HUMAN)
05-22-2500-25MGFORMYL-MET-LEU-PHE-OH
05-22-3838-1MGBQ-788, SODIUM SALT
05-22-3838-5MGBQ-788, SODIUM SALT
05-23-0050-0.1MGCORTICOTROPIN-REL. FACTOR,HUMAN+RAT
05-23-0050-1MGCORTICOTROPIN-REL. FACTOR,HUMAN+RAT
05-23-0070-1MGA-HELICAL CORTICOTROPIN-REL. FACTOR 9-41
05-23-0101-1MGANGIOTENSIN II, HUMAN
05-23-0101-25MGANGIOTENSIN II, HUMAN
05-23-0101-5MGANGIOTENSIN II, HUMAN
05-23-0111-5MG[TYR(PO3H2)4]-ANGIOTENSIN II, HUMAN
05-23-0150-1MG[ARG8]-VASOPRESSIN
05-23-0150-5MG[ARG8]-VASOPRESSIN
05-23-0300-0.5MGATRIAL NATRIURETIC FACTOR 1-28, HUMAN
05-23-0300-1MGATRIAL NATRIURETIC FACTOR 1-28, HUMAN
05-23-0310-0.5MGNATRIURETIC PEPTIDE C-TYPE HUMAN+PORCINE
05-23-0310-1MGNATRIURETIC PEPTIDE C-TYPE HUMAN+PORCINE
05-23-0500-1MGBRADYKININ
05-23-0500-25MGBRADYKININ
05-23-0500-5MGBRADYKININ
05-23-0600-1MGSUBSTANCE P
05-23-0600-25MGSUBSTANCE P
05-23-0600-5MGSUBSTANCE P
05-23-0613-1MGSENKTIDE
05-23-0701-1MGNEUROKININ A
05-23-0751-1MGA-MELANOCYTE-STIMULATING HORMONE
05-23-0751-5MGA-MELANOCYTE-STIMULATING HORMONE
05-23-0753-1MGADRENOCORTICOTROPIC HORMONE 1-24, HUMAN
05-23-0756-5MG[NLE4,D-PHE7]-A-MELANOCYTE-STIM. HORMONE
05-23-0800-1MGDYNORPHIN A 1-13, PORCINE
05-23-0850-1MGSOMATOSTATIN
05-23-0850-5MGSOMATOSTATIN
05-23-0930-1MGB-ENDORPHIN, HUMAN
05-23-1200-5MGNEUROTENSIN
05-23-1701-1MGFR-1
05-23-2005-0.5MGNEUROPEPTIDE Y, HUMAN
05-23-2005-1MGNEUROPEPTIDE Y, HUMAN
05-23-2101-0.5MGVASOACTIVE INTESTINAL PEP HUMAN+PORC+RAT
05-23-2101-1MGVASOACTIVE INTESTINAL PEP HUMAN+PORC+RAT
05-23-2102-1MGSECRETIN, HUMAN
05-23-2150-0.1MGPACAP 38, OVINE
05-23-2150-1MGPACAP 38, OVINE
05-23-2151-0.5MGPACAP 27 AMIDE, OVINE
05-23-2301-1MGGASTRIN I, HUMAN
05-23-2401-25MGCALCITONIN, SALMON
05-23-2402-0.5MGCALCITONIN, HUMAN
05-23-2402-1MGCALCITONIN, HUMAN
05-23-2404-0.5MGCALCITONIN GENE-RELATED PEPTIDE-I, HUMAN
05-23-2404-1MGCALCITONIN GENE-RELATED PEPTIDE-I, HUMAN
05-23-2540-0.5MGAMYLIN, HUMAN
05-23-2700-0.1MGGLUCAGON, HUMAN
05-23-2700-0.5MGGLUCAGON, HUMAN
05-23-3703-1MGLAMININ NONAPEPTIDE
05-23-3703-5MGLAMININ NONAPEPTIDE
05-23-3800-0.1MGENDOTHELIN 1, HUMAN AND PORCINE
05-23-3800-0.5MGENDOTHELIN 1, HUMAN AND PORCINE
05-23-3800-1MGENDOTHELIN 1, HUMAN AND PORCINE
05-23-3801-0.1MGENDOTHELIN 3, HUMAN AND RAT
05-23-3801-0.5MGENDOTHELIN 3, HUMAN AND RAT
05-23-3813-.1MGSARAFOTOXIN S6C ATRACTASPSIS ENGADDENSIS
05-23-3813-.5MGSARAFOTOXIN S6C ATRACTASPSIS ENGADDENSIS
05-23-3831-1MGBQ-123, SODIUM SALT
05-23-3831-5MGBQ-123, SODIUM SALT
05-23-4904-1MGPROTEIN KINASE C INHIBITOR PEPTIDE 19-31
05-23-4904-5MGPROTEIN KINASE C INHIBITOR PEPTIDE 19-31
05-23-4910-5MGSYNTIDE 2
05-23-5501-1MGPARATHYROID HORMONE 1-34, HUMAN
100050-5MG1400W
100064-50UG14-3-3ZETA, GST-FUSION, HU., RECOM
100065-100MG2-APB
100066-25MG5HPP-33
100068-500UG17-AAG
100069-500UG17-DMAG
100071-100UG14-3-3BETA, GST-FUSION, HUMAN, REC.
100072-100UG14-3-3GAMMA, GST-FUSION, HUMAN, REC.
100105-10MGA23187, FREE ACID, S. CHARTREUSENSIS
100105-1MGA23187, FREE ACID, S. CHARTREUSENSIS
100105-50MGA23187, FREE ACID, S. CHARTREUSENSIS
100105-5MGA23187, FREE ACID, S. CHARTREUSENSIS
100106-10MGA23187, MIXED CALCIUM-MAGNESIUM SALT
100107-1MGA23187, 4-BROMO-
100109-10MGAACOCF3
100110-1GMAAPH
100122-10MGA3, HCL
100128-5MGA77 1726
101500-100MGAEBSF, HCL
101500-1GMAEBSF, HCL
101500-500MGAEBSF, HCL
101500-50MGAEBSF, HCL
102555-2000UABL PROTEIN TYROSINE KINASE, RECOMB.
104550-25MGACA
104850-1SETACTIN POLYMERIZATION INTERFER AGENTS SET
1063-100MGACETYLCOENZYME A, TRILITHIUM SALT
1063-10MGACETYLCOENZYME A, TRILITHIUM SALT
106425-5GMN-ACETYL-L-CYSTEINE
107920-200UGANTI-ACRP30 RABBIT PAB
1079-25GMN-ACETYL-A-D-GLUCOSAMINE
108975-100MGLEUPEPTIN, HEMISULFATE
108975-10MGLEUPEPTIN, HEMISULFATE
108975-25MGLEUPEPTIN, HEMISULFATE
108975-50MGLEUPEPTIN, HEMISULFATE
108975-5MGLEUPEPTIN, HEMISULFATE
108976-10MGLEUPEPTIN, HEMISULFATE, SYNTHETIC
108976-25MGLEUPEPTIN, HEMISULFATE, SYNTHETIC
110116-1UB1-2,3,4,6-N-AC-GLUCOSAMINIDASE, RECOM
110123-5MGACETYL-11-KETO-B-BOSWELLIC ACID
110138-1GMN-ACETYLNEURAMINIC ACID, SYNTHETIC
110145-5MGD-ERYTHRO-SPHINGOSINE, N-ACETYL-
110150-5MGN-AC-D-GLUCOSAMINYL-B1,4-N-AC-MURAMYL-L-
110153-2MGN-ACETYLGALACTOSAMINE A1-O-SERINE
110175-1MGACETYL-PEPSTATIN
112251-1MGA1-ACID GLYCOPROTEIN, HUMAN SERUM
113000-1GMACRIDINE ORANGE
113200-500ULANTI-A-ACTIN MOUSE MAB (1A4)
114666-1SETACTINOMYCIN D, STREPTOMYCES SP.
114666-5MGACTINOMYCIN D, STREPTOMYCES SP.
114700-5UGACTIVIN A, HU., RECOM, CHO CELL LINE
114711-25UGFOLLISTATIN 300, HU., RECOM
114798-25MGACYCLOVIR
114802-250UGADAAHX3L3VS
114810-5MGADAMTS-5 INHIBITOR
1152-100GMADENINE
1152-10GMADENINE
1152-25GMADENINE
115500-50UGADENOPHOSTIN A, HEXASODIUM SALT
1160-100GMADENOSINE
1160-10GMADENOSINE
116530-50MGN(6),O(2\')-DIOCTANOYL-CAMP, SODIUM SALT
116801-100MGADENOSINE-3\'5\'-CYCLIC MONOPHOSPHATE, NA
116801-1GMADENOSINE-3\'5\'-CYCLIC MONOPHOSPHATE, NA
116802-10UMOLN(6)-BENZOYL-CAMP, SODIUM SALT
116805-500UGPROTEIN KINASE A INHIBITOR 5-24
116811-1KITCAMP DIRECT IMMUNOASSAY KIT
116812-10MG8-(4-CHLOROPHENYLTHIO)-CAMP, SODIUM SALT
116814-5UMOLCAMPS, RP-ISOMER, TEA SALT
116815-10UMOLCAMPS, SP-ISOMER, TEA SALT
116816-5UMOLCAMPS, 8-BROMO-, RP-ISOMER, SODIUM SALT
116818-5UMOLCAMPS, 8-BROMO-, SP-ISOMER, SODIUM SALT
116819-5UMOLADENOSINE 3,5-CYCLIC PHOSPHOTHIOATE 8-CL
116820-1STANTI-CAMP RABBIT PAB
116825-5UMOLADENOSINE 3,5-CYCLIC PHOSPHOTHIOATE,2\'-O
116833-5UMOL8-CPT-2\'-O-ME-CAMP
116850-25MGADENYLYL CYCLASE TYPE V INHIBITOR, NKY80
116850-5MGADENYLYL CYCLASE TYPE V INHIBITOR, NKY80
116880-500UADENOSINE DEAMINASE, BOVINE INTESTINE
116890-5MGADENOSINE KINASE INHIBITOR
117105-1GMADENOSINE 5\'-DIPHOSPHATE, POTASSIUM SALT
117110-1MGADP-RIBOSYLATION FACTOR 6, N-TERM., MYR
1171-1GMADENOSINE 5\'-DIPHOSPHATE, MONOSODIUM
1171-5GMADENOSINE 5\'-DIPHOSPHATE, MONOSODIUM
118110-5GMADENSOINE 5\'-MONOPHOSPHATE, DISOD. SALT
118410-10MGADENOSINE 3\'-PHOSPHATE 5\'-PHOSPHOSULFAT
118415-1SETADP-HPD, DIHYDRATE, AMMONIUM
118415-60UGADP-HPD, DIHYDRATE, AMMONIUM
118500-2MGATM KINASE INHIBITOR *KU-55933*
118501-5MGATM/ATR KINASE INHIBITOR *CGK773*
118502-2MGINSOLUTION ATM KINASE INHIBITOR
119101-50ULANTI-H+,K+-ATPASE A-SUB (1019-34) RB PAB
119102-50ULANTI-H+,K+-ATPASE, A-SUB (1-17) RB PAB
119107-1KITATP ASSAY KIT
119109-1EALUCIFERIN-LUCIFERASE
119113-5MGLUCIFERASE INHIBITOR I
119114-5MGLUCIFERASE INHIBITOR II
119117-100ULANTI-A1AR (309-326) RABBIT PAB
1191-1GMADENOSINE 5\'-TRIPHOSPHATE, DISODIUM SALT
119120-25MGATP-G-S, TETRALITHIUM SALT
119123-100ULATP-G-S, SOLUTION, TETRALITHIUM SALT
119125-1GMADENOSINE 5\'-TRIPHOSPHATE, DISODIUM
1191-25GMADENOSINE 5\'-TRIPHOSPHATE, DISODIUM SALT
119127-1SETCAGED ATP, DISODIUM SALT
119127-5MGCAGED ATP, DISODIUM SALT
119134-25MGADENOSINE 5\'-O-(2-THIODIPHOSPHATE), 3LI
1191-50GMADENOSINE 5\'-TRIPHOSPHATE, DISODIUM SALT
1191-5GMADENOSINE 5\'-TRIPHOSPHATE, DISODIUM SALT
119186-100UATP SULFURYLASE, S. CEREVISIAE
120002-25MGADENYLYL-IMIDODIPHOSPHATE, TETRALITHIUM
121000-1EAAdipogenesis Inducers Panel
121739-25GMADONITOL
121745-1MG4-AF DA
121765-2MGAG 825
121767-1MGAG 1024
12177-1KGAGAR
121800-10MGADVANCED GLYCATION ENDPRODUCT-BSA
121852-100GMAGAROSE, TYPE C
121853-100GMAGAROSE, TYPE I, MOLECULAR BIOLOGY GRADE
121853-500GMAGAROSE, TYPE I, MOLECULAR BIOLOGY GRADE
121937-2MLIMMOBILIZED ANTI-MOUSE IGG
121975-5UGW-AGATOXIN IVA, AGELENOPSIS APERTA
122117-5MGLECTIN, PEANUT, ARACHIS HYPOGAEA
122302-5UGW-AGATOXIN TK, AGELENOPSIS APERTA
123040-50MGAICA-RIBOSIDE
123041-25MGAICA-RIBOSIDE, 5\'-PHOSPHATE
123900-1MGAKTIDE-2T
124001-10TPD ANTI-AKT1 (PTHR308) RABBIT PAB
124002-100ULANTI-AKT2 RABBIT PAB
124003-1SETPD ANTI-AKT1 (PSER473) MOUSE MAB (11E6)
124005-1MGAKT INHIBITOR
124006-20UGAKT1, HIS-TAG, ACTIVATED, HUMAN, RECOMB.
124007-1KITAKT ACTIVITY IMMUNOASSAY KIT
124008-1MGAKT INHIBITOR II
124009-1MGAKT INHIBITOR III
124011-1MGAKT INHIBITOR IV
124011-5MGAKT INHIBITOR IV
124012-1MGAKT INHIBITOR V, TRICIRIBINE
124013-2MGAKT INHIBITOR VI, AKT-IN
124014-2MGAKT INHIBITOR VII, TAT-AKT-IN
124015-1MGINSOLUTION(TM) AKT INHIBITOR IV
124017-1MGINSOLUTION AKT INHIBITOR VIII
124018-1MGAKT INHIBITOR VIII, ISOZYME-SELECTIVE
124018-5MGAKT INHIBITOR VIII, ISOZYME-SELECTIVE
124019-5MGAKT INHIBITOR IX, API-59CJ-OME
124020-5MGAKT INHIBITOR X
124021-20UGAKT2, GST-FUSION PROTEIN, HU., RECOM
124022-5UGAKT3, GST-FUSION PROTEIN HU., RECOM
124023-5UGDDR2, GST-FUSION, HUMAN, REC.
124024-10UGTIE2, HIS*TAG, HUMAN, REC.
124026-50UGPCAF HISTONE ACETYL TRANSFERASE, RECOM
124027-1MGMYELIN BASIC PROTEIN, MOUSE
124028-5MGAKT INHIBITOR XI
124029-2MGAKTINHIBITORXII,ISOZYME-SELECTIVE,AKTI-2
124030-2MGAKTINHIBITORXIII,ISOZYME-SELECTIVE,AKTI2
124031-1EAINHIBITORSELECTAKT/PI 3-K/MTORSIGNALINGP
1250-100GML-ALANINE
125510-5MGDIPEPTIDYLPEPTIDASE IV SUB. I, FLUOR.
12657-1GMBSA, FRACTION V, CRYSTALLINE
12657-25GMBSA, FRACTION V, CRYSTALLINE
126575-100GMBSA, FRACTION V, FATTY ACID-FREE
126575-10GMBSA, FRACTION V, FATTY ACID-FREE
12657-5GMBSA, FRACTION V, CRYSTALLINE
126579-100GMBSA, FRAC. V, FATTY ACID-POOR, EF
126579-10GMBSA, FRAC. V, FATTY ACID-POOR, EF
126584-1MLANTI-ALBUMIN RABBIT PAB
12659-100GMALBUMIN, BOVINE SERUM, LOW HEAVY METALS
12659-1KGALBUMIN, BOVINE SERUM, LOW HEAVY METALS
12659-250GMALBUMIN, BOVINE SERUM, LOW HEAVY METALS
12659-25GMALBUMIN, BOVINE SERUM, LOW HEAVY METALS
126593-100GMBSA, FRACTION V, RIA AND ELISA GR.
126593-10GMBSA, FRACTION V, RIA AND ELISA GR.
126593-25GMBSA, FRACTION V, RIA AND ELISA GR.
12659-500GMALBUMIN, BOVINE SERUM, LOW HEAVY METALS
12660-100GMBSA, FRACTION V, MODIFIED COHN, PH 5.2
12660-1KGBSA, FRACTION V, MODIFIED COHN, PH 5.2
126609-100GMBSA, FRAC. V, FATTY ACID, NUC, PROT-FREE
126609-10GMBSA, FRAC. V, FATTY ACID, NUC, PROT-FREE
126609-5GMBSA, FRAC. V, FATTY ACID, NUC, PROT-FREE
126615-25MLBSA, 10% AQUEOUS SOLN, NUCLEASE-FREE
126621-50MLBSA, COHN FRACTION, 30% AQUEOUS SOLN
126625-50MLBSA, 30% AQUEOUS SOLN, STABILIZER-FREE
126626-50MLBSA, 30% AQUEOUS SOLN, PRESERVATIVE-FREE
126654-1GMALBUMIN, HUMAN SERUM, NON-DENATURED
126658-1GMHSA, FRACTION V, HIGH PURITY
126658-5GMHSA, FRACTION V, HIGH PURITY
12666-100GMHSA, FRACTION V, LOW HEAVY METALS
12666-10GMHSA, FRACTION V, LOW HEAVY METALS
12666-1GMHSA, FRACTION V, LOW HEAVY METALS
126674-25MGALBUMIN, MOUSE
12667-50MLHSA, 30% AQUEOUS SOLN
12668-100GMALBUMIN, HUMAN SERUM, FRACTION V
12668-10GMALBUMIN, HUMAN SERUM, FRACTION V
126705-1MLANTI-ALBUMIN RABBIT PAB
126722-100MGALBUMIN, RAT
126741-500MLALCALASE ENZYME, BACILLUS LICHENIFORMIS
126850-2.5MGA1-PDX, HUMAN, RECOMBINANT, E. COLI
126855-100MGALENDRONATE, SODIUM SALT
126870-1MGALSTERPAULLONE
126871-1MGALSTERPAULLONE, 2-CYANOETHYL
126920-25MGALDH2 AGONIST, ALDA-1
126925-100UALDEHYDE DEHYDROGENASE, YEAST
127100-10MGALLOPREGNANOLONE
128125-5MGALOISINE A
128135-5MGALOISINE, RP106
129741-1MGA-AMANITIN, AMANITA SP.

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