Identification and expression of β-defensin genes in Egyptian buffalo (Bubalus bubalis L.) infected with mastitis disease

Mastitis is a disease that causes an inflammation of mammary gland (MG). The ability of 
animal’s immune defense system to cope with infections of MG still are unknown .The β- defensin 
protein molecules of the innate system that was expressed by β- defensin gene  may be candidate 
genes for resistance to mastitis. Therefore, the present investigation was aimed to isolate these 
genes from infected lactated female animals and amplify them using PCR in the presence of specific 
β- defensin primers, examine whether mastitis activates the innate immune system or not as 
indicated by using DD-PCR technique . Results indicated that the amplified PCR products were 
2060 and 410 bp for healthy (control) and infected animals, respectively which were digested by 
Hinf1 and pvuII restriction enzymes for genotyping. Four fragments (150, 410, 600 and 900 bp) 
were produced from digestion of 2060 bp ,whereas 410 bp was digested to 180, 130 and 100 bp 
fragments that was different from healthy animals and could be associated with infection. The 
results of DD-PCR revealed several bands that were only expressed in the infected animals such as 
cDNA 2100 bp defense fragment which clearly appeared with presence of specific defense primer 
(forward).