418
Appendix
nine 12 mg, DL-valine 14 mg, glycine 2.5 mg, L-tyrosine 1 mg, L-hydroxyproline 2.5 mg,
DL-serine 12.5 mg, taurine 5 mg, ornithine dihydrochloride 5 mg, and L-asparagine 5 mg.
Adjust the pH to 7.5 with 1 N KOH and 1 N HC!.
To prepare medium C, dissolve 0.25 mg glutathione into solution C.
Wyatt's medium (Wyatt, 1956)
The concentrations are given as mg/l,OOO ml culture medium.
NaH2P04 1,100, KCl 2,980, MgCl 2 ·6H 2 0 3,040, MgS0 4 ·7H 2 0 3,700, CaCl 2 810, glucose
700, fructose 400, sucrose 400, malic acid 670, a-ketoglutaric acid 370, succinic acid 60,
fumaric acid 55, DL-a-alanine 450, ~-alanine 200, L-arginine 700, L-asparagine 350, Laspartic acid 350, cysteine 80, L-cystine 25, L-glutamic acid 600, L-glutamine 600, glycine 650, L-histidine 2,500, DL-isoleucine 100, DL-leucine 150, DL-lysine 1,250, DL-methionine 100, L-phenylalanine 150, L-proline 350, DL-serine 1,100, DL-threonine 350, Ltryptophan 100, L-tyrosine 50, and DL-valine 200. The pH of the medium is adjusted to
6.35 with 1 N KOH and 1 N HC!.
Use the medium by adding heat-treated Bombyx mori hemolymlph.
ZH 1% medium (Wyss, 1977)
Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
CaCl 2 ·2H 2 0 890, KCl 2,000, MgCl 2 ·6H 2 0 1,000, MgS0 4 ·7H 2 0 3,700, NaCl 2,050,
NaH 2 P04·2H20 470, CuS04·5HzO 0.01, FeS04·7Hp 0.6, MnS04·7H 2 0 0.3, ZnS0 4 ·7H 2 0
0.6, L-arginine 170, L-asparagine l30, L-cysteine hydrochloride 310, L-glutamine 700, Lglutamic acid 290, glycine 750, L-histidine 150, L-isoleucine l30, L-leucine l30, L-lysine
180, L-methionine 150, L-phenylalanine 160, L-proline llO, L-serine 520, L-threonine
120, L-tryptophan 100, L-tyrosine 90, L-valine 120, glucose 2,000, malic acid 670, oxaloacetic acid 250, sodium acetate·3H 2 0 25, succinic acid 60, biotin 0.01, calcium pantothenate 0.02, choline chloride 0.2, folic acid 0.02, m-inositol 0.02, niacin 0.02, niacinamide 0.02, pyridoxine hydrochloride 0.02, pyridoxal hydrochloride 0.02, riboflavin 0.02,
thiamine hydrochloride 0.02,p-aminobenzoic acid 0.02, cyanocobalamin 0.1, phenol
red 10, high molecular weight fraction of yeast extract* 1,500 (equivalent to original
yeast extract), and heat-inactivated horse serum 10 m!. The pH is adjusted to 6.6 with
NaOH.
*The fractionation of yeast extract was made using Sephadex G25 (Sigma) chromatography, and fractions 23-46 are used (Wyss, 1977).
ZW medium (Wyss, 1982)
Unless otherwise indicated, the concentrations are given as mg/1,000 ml culture medium.
L-a-Alanine 45, ~-alanine 45, L-arginine 174, L-asparagine 150, L-aspartic acid l33, Lcysteine hydrochloride 315, L-glutamine 680, L-glutamic acid 294, glycine 750, L-histidine 620, L-hydroxyproline 65, L-isoleucine l31, L-leucine l31, L-lysine hydrochloride
182, L-methionine 149, L-ornithine hydrochloride 84, L-phenylalanine 165, L-proline
115, L-serine 525, L-threonine 119, L-tryptophan 102, L-tyrosine 90, L-valine 117, glucose 2,000, inosine 2.7, thymidine 2.4, uridine 2.4, 1-thioglycerol 5.4, DL-carnitine hydrochloride 1,000, reduced glutathione 50, malic acid 670, oxaloacetic acid 250, sodium acetate·3Hp 25, sodium pyruvate llO, succinic acid 60, CaCl 2 ·2H 2 0 147, KCl
Appendix
nine 12 mg, DL-valine 14 mg, glycine 2.5 mg, L-tyrosine 1 mg, L-hydroxyproline 2.5 mg,
DL-serine 12.5 mg, taurine 5 mg, ornithine dihydrochloride 5 mg, and L-asparagine 5 mg.
Adjust the pH to 7.5 with 1 N KOH and 1 N HC!.
To prepare medium C, dissolve 0.25 mg glutathione into solution C.
Wyatt's medium (Wyatt, 1956)
The concentrations are given as mg/l,OOO ml culture medium.
NaH2P04 1,100, KCl 2,980, MgCl 2 ·6H 2 0 3,040, MgS0 4 ·7H 2 0 3,700, CaCl 2 810, glucose
700, fructose 400, sucrose 400, malic acid 670, a-ketoglutaric acid 370, succinic acid 60,
fumaric acid 55, DL-a-alanine 450, ~-alanine 200, L-arginine 700, L-asparagine 350, Laspartic acid 350, cysteine 80, L-cystine 25, L-glutamic acid 600, L-glutamine 600, glycine 650, L-histidine 2,500, DL-isoleucine 100, DL-leucine 150, DL-lysine 1,250, DL-methionine 100, L-phenylalanine 150, L-proline 350, DL-serine 1,100, DL-threonine 350, Ltryptophan 100, L-tyrosine 50, and DL-valine 200. The pH of the medium is adjusted to
6.35 with 1 N KOH and 1 N HC!.
Use the medium by adding heat-treated Bombyx mori hemolymlph.
ZH 1% medium (Wyss, 1977)
Unless otherwise indicated, the concentrations are given as mg/l,OOO ml culture medium.
CaCl 2 ·2H 2 0 890, KCl 2,000, MgCl 2 ·6H 2 0 1,000, MgS0 4 ·7H 2 0 3,700, NaCl 2,050,
NaH 2 P04·2H20 470, CuS04·5HzO 0.01, FeS04·7Hp 0.6, MnS04·7H 2 0 0.3, ZnS0 4 ·7H 2 0
0.6, L-arginine 170, L-asparagine l30, L-cysteine hydrochloride 310, L-glutamine 700, Lglutamic acid 290, glycine 750, L-histidine 150, L-isoleucine l30, L-leucine l30, L-lysine
180, L-methionine 150, L-phenylalanine 160, L-proline llO, L-serine 520, L-threonine
120, L-tryptophan 100, L-tyrosine 90, L-valine 120, glucose 2,000, malic acid 670, oxaloacetic acid 250, sodium acetate·3H 2 0 25, succinic acid 60, biotin 0.01, calcium pantothenate 0.02, choline chloride 0.2, folic acid 0.02, m-inositol 0.02, niacin 0.02, niacinamide 0.02, pyridoxine hydrochloride 0.02, pyridoxal hydrochloride 0.02, riboflavin 0.02,
thiamine hydrochloride 0.02,p-aminobenzoic acid 0.02, cyanocobalamin 0.1, phenol
red 10, high molecular weight fraction of yeast extract* 1,500 (equivalent to original
yeast extract), and heat-inactivated horse serum 10 m!. The pH is adjusted to 6.6 with
NaOH.
*The fractionation of yeast extract was made using Sephadex G25 (Sigma) chromatography, and fractions 23-46 are used (Wyss, 1977).
ZW medium (Wyss, 1982)
Unless otherwise indicated, the concentrations are given as mg/1,000 ml culture medium.
L-a-Alanine 45, ~-alanine 45, L-arginine 174, L-asparagine 150, L-aspartic acid l33, Lcysteine hydrochloride 315, L-glutamine 680, L-glutamic acid 294, glycine 750, L-histidine 620, L-hydroxyproline 65, L-isoleucine l31, L-leucine l31, L-lysine hydrochloride
182, L-methionine 149, L-ornithine hydrochloride 84, L-phenylalanine 165, L-proline
115, L-serine 525, L-threonine 119, L-tryptophan 102, L-tyrosine 90, L-valine 117, glucose 2,000, inosine 2.7, thymidine 2.4, uridine 2.4, 1-thioglycerol 5.4, DL-carnitine hydrochloride 1,000, reduced glutathione 50, malic acid 670, oxaloacetic acid 250, sodium acetate·3Hp 25, sodium pyruvate llO, succinic acid 60, CaCl 2 ·2H 2 0 147, KCl