Genetic Test for the Identification of Carriers of Complex Vertebral Malformations in Cattle

Abstract

Genetic markers for identifying bovine carriers of complex vertebral malformation (CVM) disease gene are described. The genetic markers, including the microsatellite markers BM4129, INRAA003, BMS2790, ILSTS029, INRA123, BM220, HUJ246, BMS862, BMS937, BL1048, BMS2095 and BMS1266 and the bovine SLC35A3 gene, are located on bovine chromosome BTA3. The G/T polymorphism at position 559 of the bovine SLC35A3 gene is identified as being causative and diagnostic for CVM in cattle.

1 . A method for identifying a Holstein-Fresian subject as a carrier of a genetic determinant for bovine complex vertebral malformation (CVM), the method comprising
 i) providing a sample comprising genetic material from a Holstein-Friesian subject;   ii) obtaining from said sample genomic DNA sequences from bos Taurus autosome 3 (BTA3) comprising a region between the polymorphic microsatellite markers INRAA003 and BMS937;   iii) determining the sequence of at least a part of said region;   iv) determining whether said sequence comprises a sequence polymorphism that is genetically linked to the SLC35A3 gene polymorphism having a T nucleotide at a position equivalent to position 559 of SEQ ID NO:17; and   v) identifying said bovine Holstein-Friesian subject as a carrier of a genetic determinant for CVM, when a sequence polymorphism linked to the SLC35A3 gene polymorphism having a T nucleotide at a position equivalent to position 559 of SEQ ID NO: 17 is detected in step iv).   
     
     
         2 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is located in the region flanked by and including the polymorphic microsatellite markers INRAA003 and ILSTS029. 
     
     
         3 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker INRAA003. 
     
     
         4 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker BMS2790. 
     
     
         5 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker ILSTS029. 
     
     
         6 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker INRA123. 
     
     
         7 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker BM220. 
     
     
         8 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker HUJ246. 
     
     
         9 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker BMS862. 
     
     
         10 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism is the microsatellite marker BMS937. 
     
     
         11 . The method according to  claim 1 , wherein the sequence of at least a part of said region between the polymorphic microsatellite markers INRAA003 and BMS937 is determined by a technique selected from the group consisting of allele-specific PCR, minisequencing, primer extension, pyro-sequencing, PCR-RFLP, allele-specific rolling circle amplification, and primer extension followed by MALDI-TOF mass spectrometry. 
     
     
         12 . The method according to  claim 1 , wherein the genetically linked sequence polymorphism has a LOD score of at least 3.0 with respect to the SLC35A3 gene polymorphism having a T nucleotide at a position equivalent to position 559 of SEQ ID NO: 17.