FreshPatents.com Logo
stats FreshPatents Stats
n/a views for this patent on FreshPatents.com
Updated: October 13 2014
newTOP 200 Companies filing patents this week


    Free Services  

  • MONITOR KEYWORDS
  • Enter keywords & we'll notify you when a new patent matches your request (weekly update).

  • ORGANIZER
  • Save & organize patents so you can view them later.

  • RSS rss
  • Create custom RSS feeds. Track keywords without receiving email.

  • ARCHIVE
  • View the last few months of your Keyword emails.

  • COMPANY DIRECTORY
  • Patents sorted by company.

Follow us on Twitter
twitter icon@FreshPatents

Oligonucleotide sequences that identify species of animal

last patentdownload pdfdownload imgimage previewnext patent


20120270223 patent thumbnailZoom

Oligonucleotide sequences that identify species of animal


The present invention provides a method for identifying animal species, said method comprises a step of amplifying a DNA fragment by PCR using a DNA in a sample as a template and animal-specific DNA sequences as a primer pair, wherein the animal-specific DNA sequences are derived from a ATP synthase subunit 8 gene or a region proximal thereto of a mitochondrial genome; and a step of detecting the amplified DNA fragment.
Related Terms: Dna Sequences

Browse recent National Institute Of Agrobiological Sciences patents - Tsukuba-shi, JP
Inventors: Toyoko Kusama, Koichi Kadowaki, Tetsuya Nomura
USPTO Applicaton #: #20120270223 - Class: 435 612 (USPTO) - 10/25/12 - Class 435 


view organizer monitor keywords


The Patent Description & Claims data below is from USPTO Patent Application 20120270223, Oligonucleotide sequences that identify species of animal.

last patentpdficondownload pdfimage previewnext patent

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a Division of application Serial No. 12/707,828 filed Feb. 18, 2010, which is a Continuation of application Ser. No. 10/826,119 filed Apr. 16, 2004,the entire contents of both of which is hereby incorporated by reference. Applicants request use of the compliant computer readable “Sequence Listing” that is already on file for application Ser. No. 10/826,119 filed Apr. 16, 2004 and state that the paper or compact disc copy of the “Sequence Listing” in this application is identical to the computer readable copy filed for application Ser. No. 10/826,119 filed Apr. 16, 2004.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to methods for identifying animal species and animal-derived DNA-specific primer pairs used therein. More specifically, the present invention relates to methods for identifying animal species that include a step of amplifying an animal-specific DNA sequence derived from the ATP synthase subunit 8 gene of a mitochondrial genome, and primer pairs used in this amplification step.

2. Description of the Related Art

Currently, there is a problem that cattle is infected with bovine spongiform encephalopathy (BSE) by giving feed containing meat and bonemeal derived from cattle infected with BSE. It has been shown that BSE-like diseases may be present in various livestock as well. Accordingly, since the emergence of BSE, there has been a need to develop sensitive and effective method to identify feed contaminated bone and bonemeal, and this has become a particularly urgent matter for authorities.

Immunological methods and gene identification methods using nuclear gene conventionally have been used as methods for identifying animal species. Examples of immunological methods include ELISA and immunoblotting. PCR is an example of a gene identification method using nuclear gene. However, there are many problems with methods for identifying the animal species that are currently employed. For example, in meat and bonemeal that has been heat-treated, there is a high likelihood that nucleic acids have been fragmented. Furthermore, majority of feed in which meat and bonemeal has been mixed is composed of plant-derived material. Therefore, it is necessary to analyze trace amounts of animal-derived components. There is a dire need for the development of a detection method that is highly sensitive and effective and that can be executed with respect to such heat-treated samples.

Accordingly, a method other than immunological methods and gene identification methods employing nuclear gene that is for detecting and identifying the animal species of animal-derived components that are present in trace amounts is desirable. In particular, it is crucial to identify the type of animal meat, meat and bonemeal, or fishmeal used in feed given to livestock and pets. Furthermore, it is desirable that the detection method is highly sensitive and differentiate species of animal DNA present in trace amounts from among large quantities of plant DNA or DNA of other animal species.

SUMMARY

OF THE INVENTION

The inventors of the present application focused on the mitochondrial genome, which is inherited maternally and exists in a greater number of copies than nuclear genome, and investigated the use of mitochondrial gene as a target for identifying animal species. Their research indicated that the homologous sequence with ATP synthase subunit 8 gene (atp8 gene) from animal mitochondrial genome is not present in plant (Oryza sativa) mitochondrial genome. Thus, the inventors found that the DNA derived from the atp8 gene can serve as a material for specific detection of trace amounts of animal DNA among plant-based feed, in other word, plant atp8 gene is very diverged from animal atp8 gene, and also that specific DNA sequences of the animal mitochondrial atp8 gene can be used to identify the animal species, thereby arriving at the present invention.

The present invention provides a method for identifying animal species, said method comprises:

amplifying a DNA sequence by PCR using a DNA in a sample as a template and animal-specific DNA sequences as a primer pair, wherein the animal-specific DNA sequences are derived from a ATP synthase subunit 8 gene or a region proximal thereto of a mitochondrial genome, and

detecting the amplified DNA sequence.

In a preferred embodiment, the animal is a mammal, and further preferably, the primer pair is a combination of the DNA sequence of SEQ ID NO: 1 and the DNA sequence of SEQ ID NO: 2.

In a further preferred embodiment, the mammal is selected from the group consisting of cattle, sheep, goat, deer, pig, horse, rabbit, and whale.

In a preferred embodiment, the animal is a ruminant, and further preferably, the primer pair is a combination of the DNA sequence of SEQ ID NO: 3 and the DNA sequence of SEQ ID NO: 4, or a combination of the DNA sequence of SEQ ID NO: 5 and the DNA sequence of SEQ ID NO: 6.

In a further preferred embodiment, the ruminant is selected from the group consisting of cattle, sheep, goat, and deer.

In a preferred embodiment, the animal is a cattle, and further preferably, the primer pair is a combination of DNA sequences selected from the group consisting of the following DNA sequence combinations: SEQ ID NO: 9 and SEQ ID NO: 13; SEQ ID NO: 9 and SEQ ID NO: 12; SEQ ID NO: 11 and SEQ ID NO: 13; SEQ ID NO: 10 and SEQ ID NO: 12; SEQ ID NO: 11 and SEQ ID NO: 12; SEQ ID NO: 8 and SEQ ID NO: 12; and SEQ ID NO: 14 and SEQ ID NO: 15.

In a preferred embodiment, the animal is a pig, and further preferably, the primer pair is a combination of the DNA sequence of SEQ ID NO: 17 and the DNA sequence of SEQ ID NO: 19, or a combination of the DNA sequence of SEQ ID. NO: 18 and the DNA sequence of SEQ ID NO: 22.

In a preferred embodiment, the animal is a sheep, and further preferably, the primer pair is a combination of the DNA sequence of SEQ ID NO: 23 and the DNA sequence of SEQ ID NO: 24.

In a preferred embodiment, the animal is a goat, and further preferably, the primer pair is a combination of the DNA sequence of SEQ ID NO: 25 and the DNA sequence of SEQ ID NO: 26.

In a preferred embodiment, the animal is a chicken, and further preferably, the primer pair is a combination of the DNA sequence of SEQ ID NO: 28 and the DNA sequence of SEQ ID NO: 30.

In a preferred embodiment, the animal is a fish, and further preferably, the primer pair is a combination of the DNA sequence selected from the group consisting of SEQ ID NOS: 32, 34, 38 and 39 and the DNA sequence selected from the group consisting of SEQ ID NOS: 33, 35, 36, 37, 40, and 41.

In a further preferred embodiment, the fish is selected from the group consisting of sardine, flatfish, salmon, Alaska Pollack, tuna, and lady crab.

In a further preferred embodiment, the sample is selected from a group consisting of raw meat, raw fish, processed meat food products, processed fish food products, food products containing processed meat, food products containing processed fish, blood, hair, body fluids, milk, milk processing products, meat and bonemeal, bonemeal, fishmeal, fish soluble, and feed, fertilizer, and feed additive containing them.

The present invention also provides a primer pair for detection of a mammal-specific DNA, the primer pair being a combination of the DNA sequence of SEQ ID NO: 1 and the DNA sequence of SEQ ID NO: 2.

In a preferred embodiment, the mammal is selected from the group consisting of cattle, sheep, goat, deer, pig, horse, rabbit, and whale.

The present invention also provides a primer pair for detection of a ruminant-specific DNA, the primer pair being a combination of the DNA sequence of SEQ ID NO: 3 and the DNA sequence of SEQ ID NO: 4, or a combination of the DNA sequence of SEQ ID NO: 5 and the DNA sequence of SEQ ID NO: 6.

In a preferred embodiment, the ruminant is selected from the group consisting of cattle, sheep, goat, and deer.

The present invention also provides a primer pair for detection of a cattle-specific DNA, the primer pair being a combination of DNA sequences selected from the group consisting of the, following DNA sequence combinations: SEQ ID NO: 9 and SEQ ID NO: 13; SEQ ID NO: 9 and SEQ ID NO: 12; SEQ ID NO: 11 and SEQ ID NO: 13; SEQ ID NO: 10 and SEQ ID NO: 12; SEQ ID NO: 11 and SEQ ID NO: 12; SEQ ID NO: 8 and SEQ ID NO: 12; and SEQ ID NO: 14 and SEQ ID NO: 15.

The present invention also provides a primer pair for detection of a pig-specific DNA, the primer pair being a combination of the DNA sequence of SEQ ID NO: 17 and the DNA sequence of SEQ ID NO: 19, or a combination the DNA sequence of SEQ ID NO: 18 and the DNA sequence of SEQ ID NO: 22.

The present invention also provides a primer pair for detection of a sheep-specific DNA, the primer pair being a combination of the DNA sequence of SEQ ID NO: 23 and the DNA sequence of SEQ ID NO: 24.

The present invention also provides a primer pair for detection of a goat-specific DNA, the primer pair being a combination of the DNA sequence of SEQ ID NO: 25 and the DNA sequence of SEQ ID NO:\'26.

The present invention also provides a primer pair for detection of a chicken-specific DNA, the primer pair being a combination of the DNA sequence of SEQ ID NO: 28 and the DNA sequence of SEQ ID NO: 30.

The present invention also provides a primer pair for detection of a fish-specific DNA, the primer pair being a combination of the DNA sequence selected from the group consisting of SEQ ID NOS: 32, 34, 38 and 39 and the DNA sequence selected from the group consisting of SEQ ID NOS: 33, 35, 36, 37, 40, and 41.

In a preferred embodiment, the fish is selected from the group consisting of sardine, flatfish, salmon, Alaska Pollack, tuna, and lady crab.

The present invention also provides a primer pair for detection of a plant-specific DNA, the primer pair being a combination of the DNA sequence of SEQ ID NO: 42 and the DNA sequence of SEQ ID NO: 43.

Further, the present invention provides a method for detecting animal-derived components present in mixed feed, said method comprises:

amplifying a DNA sequence by PCR using a DNA in a sample as a template and animal-specific DNA sequences as a primer pair, wherein the animal-specific DNA sequences are derived from a ATP synthase subunit 8 gene or a region proximal thereto of a mitochondrial genome, and

detecting the amplified DNA sequence.

The present invention also provides a kit for detecting an animal-derived component present in a sample, said kit comprises at least one of primer pair specific for any animal described above.

In a preferred embodiment, the kit further comprises a primer pair for detection of a plant-specific DNA, said primer pair is a combination of the DNA sequence of SEQ ID NO: 42 and the DNA sequence of SEQ ID NO: 43.

Further, the present invention provides a method for detecting plant-derived components present in sample, said method comprises:

amplifying a DNA sequence by PCR using a DNA in a sample as a template and plant-specific DNA sequences as a primer pair, wherein the plant-specific DNA sequences are derived from a ATP synthase subunit 8 gene or a region proximal thereto of a mitochondrial genome, and

detecting the amplified DNA sequence.

Using the method of the present invention, it is possible to detect, with high sensitivity, trace amounts of animal-derived DNA in a sample. It is thus applicable to identify the animal species of trace amounts of meat and bonemeal mixed into feed. More particularly, detection is possible even if trace amounts of cattle-derived meat and bonemeal are mixed into mixed feed for livestock. Further, the primers for the detection of fish-specific DNA according to the present invention are useful for detecting a wide range of fishmeal derived from various fish species that has been mixed into mixed feed or the like.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows a sequence alignment of mitochondrial atp8 genes derived from various types of animals.

FIG. 2 shows a sequence alignment of chicken, cattle, and pig mitochondrial atp8 genes.

FIG. 3 shows a sequence alignment of mitochondrial atp8 genes, and nearby regions, derived from various types of animals.

FIG. 4 shows the positional relationship on the mitochondrial atp8 gene of the primers used to specifically detect cattle-derived DNA sequences.

FIG. 5 is a photograph showing the results of electrophoresis after performing PCR using a primer pair (anicon5 and anicon3) for detecting mammal-specific DNA sequences and with DNAs derived from various types of animals serving as templates.



Download full PDF for full patent description/claims.

Advertise on FreshPatents.com - Rates & Info


You can also Monitor Keywords and Search for tracking patents relating to this Oligonucleotide sequences that identify species of animal patent application.
###
monitor keywords



Keyword Monitor How KEYWORD MONITOR works... a FREE service from FreshPatents
1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored.
3. Each week you receive an email with patent applications related to your keywords.  
Start now! - Receive info on patent apps like Oligonucleotide sequences that identify species of animal or other areas of interest.
###


Previous Patent Application:
Methods of sequencing fluorophore-quencher fret-aptamers
Next Patent Application:
Predictors of patient response to treatment with egf receptor inhibitors
Industry Class:
Chemistry: molecular biology and microbiology
Thank you for viewing the Oligonucleotide sequences that identify species of animal patent info.
- - - Apple patents, Boeing patents, Google patents, IBM patents, Jabil patents, Coca Cola patents, Motorola patents

Results in 0.69469 seconds


Other interesting Freshpatents.com categories:
Amazon , Microsoft , IBM , Boeing Facebook

###

Data source: patent applications published in the public domain by the United States Patent and Trademark Office (USPTO). Information published here is for research/educational purposes only. FreshPatents is not affiliated with the USPTO, assignee companies, inventors, law firms or other assignees. Patent applications, documents and images may contain trademarks of the respective companies/authors. FreshPatents is not responsible for the accuracy, validity or otherwise contents of these public document patent application filings. When possible a complete PDF is provided, however, in some cases the presented document/images is an abstract or sampling of the full patent application for display purposes. FreshPatents.com Terms/Support
-g2--0.7477
     SHARE
  
           

FreshNews promo


stats Patent Info
Application #
US 20120270223 A1
Publish Date
10/25/2012
Document #
File Date
10/21/2014
USPTO Class
Other USPTO Classes
International Class
/
Drawings
0


Dna Sequences


Follow us on Twitter
twitter icon@FreshPatents