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Methods for gamete production in birds

A technology for poultry and sperm, applied in biochemical equipment and methods, microorganisms, poultry industry, etc., can solve problems such as insufficient identification of chimeras

Inactive Publication Date: 2003-05-21
NORTH CAROLINA STATE UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Morphology alone is insufficient to identify mosaicism, so other markers must be supplemented

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1-8

[0023] Materials and methods

example 1

[0025] Plasmid isolation and verification

[0026] The transformed DH5α cells provided by Dr. M. Matzke were streaked onto LB plates containing the antibiotics ampicillin (20 μg / ml) + methicillin (80 μg / ml), and grown at 37° C. overnight. Six independent colonies were picked and grown overnight in 10 ml LB containing the above antibiotics. Plasmid DNA was isolated from the above 6 different colonies using the Qiagen miniprep method. To verify the identity of the plasmids, undigested, linearized (EcoRI) and double digested (EcoRI+HindIII) plasmids were separated on a 2% agarose gel. Two of the six colonies containing the insert were subsequently used for large-scale plasmid isolation (Qiagen). Separation of undigested parental plasmid (PUC18), undigested recombinant plasmid, linearized plasmid (EcoRI / HindIII / BamHI) and double digested plasmid (EcoRI+HindIII and EcoRI+BamHI) on a 2% gel , in order to confirm the identity of the isolated plasmid.

example 2

[0028] PCR labeled TM1 probe

[0029] A pair of primers were synthesized based on their ability to amplify the insert at the multiple cloning site of the parental pUC18 plasmid. These primers were M13puc reverse=5'AAC AGC TAT GAC CAT G and M13puc forward=5'GTA AAA CGA CGG CCA GT. The optimized PCR mix consisted of 3 mM MgCl in Taq buffer (Idaho Tech), 0.5 μM of each primer, 50 ng of DNA (TM1) circular denatured plasmid, 5 units of Taq polymerase (Promega), 10 μl of PCR dig-labeling mixture (Boehringer Mannheim). The volume of the reaction was adjusted to 100 microliters with sterile water, and the PCR conditions included initial denaturation at 96°C for 5 minutes, followed by 30 rounds of the following steps: denaturation (94°C) for 45 seconds, annealing (50°C) for 55 seconds, It was then extended at 72°C for 60 seconds. The PCR was performed on a "Minicycler" model PTC150 (MJ Research, MA). After amplification, all samples were electrophoresed on 2% gels...

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PUM

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Abstract

A method for the production and collection of avian sperm comprises the steps of: providing primordial germ cells from a donor avian species; administering the primordial germ cells to a recipient avian species in ovo; incubating the recipient avian species to hatch; and then collecting sperm of the donor avian species from the recipient avian species. For example, the donor avian species may be a whooping crane, and the recipient avian species may be a sand hill crane. In another example, the donor avian species may be a turkey, and the recipient avian species may be a chicken.

Description

field of invention [0001] The present invention relates to a method of transferring primordial germ cells into an avian in order to produce gametes from said avian. The method can be used for the conservation of endangered avian species, shortening the time required to produce sperm in slow-growing species such as turkey, reducing the cost of maintaining a breeder population, and altering the sex ratio of offspring populations (e.g., increasing production efficiency) . Background of the invention [0002] The ability to more conveniently produce gametes of a particular avian species is extremely useful for avian medicine and poultry production. For endangered species such as whooping cranes, the easy availability of male sperm is extremely useful. For commercial poultry such as turkeys, it is desirable to produce male sperm more quickly and economically. For meat producing poultry, there is a need for a method of increasing the proportion of male bi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K45/00A01K67/02C12N5/07C12N5/073C12N5/076
CPCA01K45/007A01K67/02A01K45/00
Inventor S·L·帕迪J·N·佩蒂特S·德科斯塔
Owner NORTH CAROLINA STATE UNIV
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