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Method of detecting sexual differentiation disruptor

A gender, female technology, applied in biochemical equipment and methods, microbial determination/inspection, animal/human peptides, etc., can solve problems such as long time, measurement skills, and difficulty in testing samples

Inactive Publication Date: 2005-08-10
TAKARA HOLDINGS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, the determination also requires skill
Also, it is difficult to handle a large number of test samples with this method
As mentioned above, in vivo analysis requires a long time from the start of evaluation, and it is difficult to analyze a large number of test samples at a time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0073] The following examples describe the present invention in further detail, but should not be interpreted as

[0074] Limitation of Scope of Invention.

example 1

[0075] Example 1: Identification of female-specific genes by subtractive crosses

[0076] One male and one female were bred in 3 liters of green water (purified water containing Chlorella chlorella) at 25°C under a 14-hour light cycle and a 10-hour dark cycle. on mature Qurt medaka (Zoological Sciences, 15:123-126, 1998). Feed 3-5 times a day with TetraMin, set the amount of TetraMin so that it is eaten within 3-5 minutes and then ovulates. On day 4 after ovulation, embryos were genotyped into males and females by examining the expression of yellow pigment genes based on autofluorescence under a fluorescent microscope. mRNA was prepared from samples from 4 periods (37 / 39 period (2-3 days before hatch), or 1, 5 or 30 days after hatch) classified according to the Iwamatsu classification table (Iwamatsu, T. Zoology Science, 11:825-839, 1994). cDNA was prepared using oligo(dT) primers and Copy kit (Invitrogen). The cDNA was cleaved with the restriction enzyme AluI, ligated wit...

example 2

[0079] Example 2: Detection of sex differentiation blocking activity of 17β-estradiol

[0080] One male and one female were bred in 3 liters of green water (purified water containing Chlorella chlorella) at 25°C under a 14-hour light cycle and a 10-hour dark cycle. on mature Qurt medaka (Zoological Sciences, 15:123-126, 1998). Feed 3-5 times a day with TetraMin, set the amount of TetraMin so that it is eaten within 3-5 minutes and then ovulates. After ovulation, eggs from 5 pairs were placed in Petri dishes, each egg was separated with forceps in dechlorinated tap water, and washed. The above water was replaced with a 1-ppb 17β-estradiol (E2) aqueous solution containing 0.1% dimethyl sulfoxide. This mixture was dispensed into wells of a 96-well round bottom microplate (#3797, Corning) rinsed thoroughly with ultrapure water so that each well contained one egg. Cover with a lid and place the microplate in an incubator at 25°C. On the 3rd day after the initiation of the cultu...

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PUM

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Abstract

Killifish genes which are expressed specifically to females in accordance with the phenotype sex, characterized by having a base sequence selected from among the base sequences represented by SEQ ID NOS:1 to 21 in Sequence Listing.

Description

technical field [0001] The present invention relates to a method for rapidly evaluating the endocrine-blocking activity of chemical substances, and a series of related technologies in the fields of medicine, pharmacology, environmental research and nutrition. Background technique [0002] Endocrine blocking agents (commonly referred to as environmental hormones) generally refer to chemicals released into the environment that have been shown to have hormone-like or antihormonal activity. It has been reported that altered reproductive capacity (especially male-to-female conversion), reduced reproductive capacity, reduced hatchability, reduced offspring survival, and abnormal reproductive behavior are due to endocrine obstruction. Caused by disconnecting agents affecting wildlife ecosystems. It has been speculated that the decrease in sperm count, endometriosis, infertility, ovarian cancer, uterine cancer, and prostate cancer are caused by the impact of endocrine blocking agen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/46
CPCC07K14/461C12N15/11
Inventor 金森章木下政人高嶋良吉蝶野英人水谷滋利近藤昭宏加藤郁之进
Owner TAKARA HOLDINGS
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