Provided is a method for evaluating immunotoxicity of a chemical substance to a mammal, including step 1: bringing a chemical substance into contact with a mammalian cell for immunotoxicity evaluation, the mammalian cell having a first reporter gene under control of a constant expression promoter and a second reporter gene under control of a promoter for immunotoxicity evaluation incorporated in a transiently or stably expressible manner; and step 2: evaluating an increase or decrease in expression of the second reporter gene relative to that of the first reporter gene and an increase or decrease in expression of the first reporter gene in the presence and absence of the chemical substance.

The present invention provides a method, of producing a useful substance, including secreting and producing a useful substance in broth by yeast contained in the broth, the yeast being a Saccharomyces strain, the broth containing a compound (A) represented by a formula (1), the compound (A) having an HLB value of 0.1 to 16 and a number average molecular weight of 200 to 30000, the weight percentage of the compound (A) based on the weight of the broth being 0.0001 to 10 wt %:

HO(A.sup.1O)m.sup.1-(A.sup.2O)m.sup.2-(A.sup.3O) m.sup.3H (1)

wherein A.sup.1O, A.sup.2O, and A.sup.3O each independently represent a C2-C4 oxyalkylene group; A.sup.1O and A.sup.2O have different structural formulas, A.sup.2O and A.sup.3O have different structural formulas;
and m.sup.1, m.sup.2, and m.sup.3 each respectively represent the average number of moles of A.sup.1O, A.sup.2O, and A.sup.3O added, and are each independently a number in the range of 1 to 600.

Luciferase polypeptides with improved light-emitting activity and their encoding nucleic acids are disclosed. These molecules are useful in a range of assays including luciferase-based gene reporter assays, bioluminescence resonance energy transfer assays, protein complementation assays and other applications in which luciferase enzymes are utilized as detectable and/or quantifiable labels. Methods and compositions for increasing the sensitivity and/or improving the kinetics of luciferase-catalyzed reactions as well as decreasing the impact of undesirable variables are also disclosed.

The screening method of the present invention is useful for screening drugs such as insulin secretagogues having an insulin secretagogue activity with minimized side effects (hypoglycemia induction, etc.). The transformant in which a polynucleotide encoding the fusion protein used for the screening method is introduced, the screening kit comprising the transformant, etc. are also useful for screening excellent drugs.

A novel luciferase that is distinct from conventional luciferase has been desired. A luciferase mutant comprising an amino acid sequence in which glutamic acid at position 4, arginine at position 11, leucine at position 18 and valine at position 27 are substituted with other amino acids, in the amino acid sequence of SEQ ID NO: 2.

A novel luciferase that is distinct from conventional luciferase has been desired. A luciferase mutant comprising an amino acid sequence in which glutamic acid at position 4, arginine at position 11, leucine at position 18 and valine at position 27 are substituted with other amino acids, in the amino acid sequence of SEQ ID NO: 2.

The present invention provides a method of designing an optimized gene which comprises altering a nucleotide sequence of a target protein gene, so that only preferential codons with high frequency of use in human cells are selected and a GC content of not less than 60% is achieved. A gene design method which involves the feature only preferential codons with high frequency of use are selected and a GC content of not less than 60% is achieved can be established as a general rule for preparing proteins with high expression level, in order to obtain chemically synthesized genes for proteins capable of high-level expression in eukaryotes.

The present invention provides a luminescent enzyme protein comprising the amino acid sequence represented by SEQ ID NO: 2, and a mutant enzyme thereof.

The invention relates to a bioluminescent substrate suitably usable in a series of artificial luciferases (ALuc), and the invention provides a wavelength-shifted spectrum with a selective high intensity luminescence and high luminescence stability obtained by the use of the substrate together with ALuc. The luminescent substrate for ALuc obtained by the invention can be included together with a suitable luminescence solution in a luminescence kit. The bioluminescent substrate for ALuc of the invention can exhibit unprecedented excellent luminescence specificity and functionality in the conventional bioluminescence probe, two-hybrid assay, bioluminescent capsule, and reporter gene assay.

The present invention relates to compositions, methods and kits using cell phenotype altering polynucleotides, cell phenotype altering primary transcripts and cell phenotype altering mmRNA molecules.