Prolactin receptor gene as a genetic marker for increased litter size in animals

a technology of prolactin receptor and gene, which is applied in the field of genetic differences for reproductive efficiency among pigs, can solve the problems of low heritability of litter size, major limiting factor in the efficient production of pork, and reproductive efficiency,

Inactive Publication Date: 2006-05-11
IOWA STATE UNIV RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0023] The invention further comprises a kit for evaluating a sample of pig DNA for the presence in pig genetic material of a desired genetic marker located in the pig prolactin receptor gene indicative of the inheritable trait of large litter size. At a minimum, the kit is a container with one or more reagents that identify a polymorphism in the pig prolactin receptor gene. Preferably, the reagent is a set of oligonucleotide primers capable of amplifying a fragment of the pig prolactin receptor gene that contains the polymorphism. Preferably, the kit further contains a restriction enzyme that cleaves the pig prolactin receptor gene in at least one place.

Problems solved by technology

Reproductive efficiency, which can be defined as the number of pigs produced per breeding female, is the major limiting factor in the efficient production of pork.
Heritability for litter size is low (10%-15%), and standard genetic methods of selecting breeding females on the basis of past litter size have not been effective.
However, it has been demonstrated that the short form is not capable of activating transcription of the milk protein genes.

Method used

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  • Prolactin receptor gene as a genetic marker for increased litter size in animals
  • Prolactin receptor gene as a genetic marker for increased litter size in animals
  • Prolactin receptor gene as a genetic marker for increased litter size in animals

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0083] Due to their high sequence homology and similarity in transcript processing, human (Boutin et al. 1989) and rabbit (Edery et al. 1989) cDNA sequences encoding the prolactin receptor were used to design degenerate primers overlapping the 31 coding and untranslated region. The primers amplified a fragment of approximately 500 base pairs in pig genomic DNA samples and human control. The forward primer 5′-TCA CAA GGT CAA C / TAA AGA TG-3′ (SEQ ID NO:4) and the reverse primer 5′-TGG / A AGA AAG / A AGG CAA G / ATG GT-3′ (SEQ ID NO:5)were used in the following PCR conditions: 93° C. for 3 minutes, 6 cycles of 93° C. 30 seconds, 47° C. 2 minutes, 72° C. 3 minutes, 36 cycles of 93° C. 30 seconds, 53° C. 2 minutes, 72° C. 5 minutes, and a final 72° C. 5 minutes. The Taq polymerase was added last while samples were held at 80° C.

[0084] Fragments from two animals were purified and sequenced in forward and reverse directions. The pig sequence from the coding region was translated to amino acids...

example 2

[0085] PCR TEST for Prolactin Receptor Genetic Marker The PCR amplification test was optimized with the following parameters.

[0086] Primers:

(SEQ ID NO:1)forward primer 5′-CCCAAAACAGCAGGAGAACG-3′(SEQ ID NO:2)reverse primer 5′-GGCAAGTGGTTGAAAATGGA-3′

[0087] PCR Conditions:

Cocktail Mix25uL reaction10X PCR buffer (Promega)2.5uL25 mM MgCl2 (Promega)2.0uL10 mM dNTP's (Boehringer Mannheim)0.5uL20 pmol / uL forward primer0.5uL20 pmol / uL reverse primer0.5uLdd Sterile H2O17.5uL12.5 ng / uL DNA1.5uLTaq Polymerase (Promega)0.125uL

[0088] The first six reagents should be mixed and 18.5 uL of this pre-mix added to each reaction tube. Add the DNA next and then overlay with a drop of sterile mineral oil. Place the reaction tubes on the terminal cycler held at 80° C. Mix the Taq with the remaining cocktail and add 5 uL to the reaction tubes, making sure to submerge the tip beneath the oil.

Thermal Cycler Program:

[0089] 1. 93° C. 3 minutes [0090] 2. 93° C. 30 seconds [0091] 3. 60° C. 1 minute [0092]...

example 3

Association of Genotype with Litter Size

[0101] The PCR test was run as detailed in Example 2 on several sows from Pig Improvement Company, (PIC) . The animals used were PIC line 19 sows which farrowed within a six month period and gilts which were born during this period that would be kept as breeding stock. Blood or tissue samples were collected and shipped to the laboratory where the DNA was extracted and used in the PRLR PCR test. Females had one to three records used in the analysis. Estimated Breeding Value for Total Number Born (BV TNB) was estimated using a mixed linear model where each successive parity of the sow is treated as a repeated record. Only the first three parities of a sow were used. The model includes the covariate of age at farrowing nested within parity, fixed effects of parity, service type (natural or AI), farm-month farrowed, and random permanent environmental and animal effects. Current h2 is assumed as 0.10 and repeatability as 0.21. Average Number Born ...

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Abstract

Disclosed herein are genetic markers for animal litter size, methods for identifying such markers, and methods of screening animals to determine those more likely to produce larger litters and preferably selecting those animals for future breeding purposes. The markers are based upon the presence or absence of certain polymorphisms in the prolactin receptor gene.

Description

CROSS REFERENCE TO RELATED APPLICATION [0001] This application is a divisional of co-pending commonly owned U.S. patent application Ser. No. 09 / 900,063 filed Jul. 6, 2001, which is a continuation-in-part of co-pending commonly owned U.S. patent application Ser. No. 09 / 274,655 filed Mar. 23, 1999, which is a continuation of co-pending, commonly owned U.S. patent application Ser. No. 08 / 812,208 filed Mar. 6, 1997 which is a continuation of co-pending, commonly owned U.S. patent application Ser. No. 08 / 742,805 filed Nov. 1, 1996, now abandoned, which is a continuation of co-pending commonly owned U.S. provisional application Ser. No. 60 / 022,180 filed Jul. 19, 1996, entitled DNA POLYMORPHISMS IN GENES THAT ARE USEFUL FOR TESTING AND SELECTING FOR INCREASED LITTER SIZE IN PIGS, priority is claimed under 35 U.S.C. Section 120.GRANT REFERENCE [0002] Work for this invention was funded in part by a grant from the United States Department of Agriculture, IAHEES / Hatch IOWO3148. The Government ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/68C07H21/04C07K14/705C07K14/72
CPCA01K2267/02C07K14/70542C07K14/70567C07K14/72C12Q1/683C12Q1/6876C12Q2600/124
Inventor ROTHSCHILD, MAXVINCENT, AMYTUGGLE, CHRISTOPHERGLADNEY, CHRISTYMILEHAM, ALANSOUTHWOOD, OLWENPLASTOW, GRAHAMSARGENT, CAROLE
Owner IOWA STATE UNIV RES FOUND
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