Investigation of DNaseIIβ Transcripts X1 and X2 in Chickens

Jordan LeSage, Joshua Grullon, and Kaylee Kidd

Abstract:
In mammals, the specific gene DNase2α is a key factor in survival after birth. DNase2α is responsible for breaking down the DNA that accumulates in the organism after a cell has died. In mice studies, loss of the DNase2α enzyme led to complications including death in utero, or shortly after birth. This finding confirms that the enzyme is essential for the life of mammals. However, the DNase2α gene is not present in the genetic makeup of chickens. So how are they surviving if DNase2α is essential? In addition to DNase2α, there is a second DNase2 gene called DNase2β. Our working hypothesis is that DNase2β, which is found in chickens, will compensate for the loss of DNase2α. Recently, two new chicken DNase2β transcripts called X1 and X2 have been found. We are confirming the presence of X1 and X2 in chicken lung and intestine cells in order to determine if these two transcripts may exhibit DNase2α activity. Over the past few weeks, the RNA in these tissues was reverse transcribed, turning the RNA into cDNA. Using this cDNA, as well as X1 and X2 specific primers, we performed PCR to find and amplify any X1 and X2 transcripts. Gel electrophoresis was performed to visualize the X1 and X2 PCR products.