Genetic Background Studying On Atp11C Gene-Trap Mice

As a lipid flip enzyme,Atp11c plays an important role in the metabolism,immune and other life activities. Atpllc transporter is the key role to lymphoid B cell differentiation and bile transport in vivo. Atp11c defects can cause symptoms such as body weight decreasing and dystocia in female mice, which suggests that Atpllc regulation may affect the growth and development,yet there are no enough evidences to prove that so far.However, this problem can probably be solved by establishing Atp11c defect mice models.Gene trapping is the most time and cost-effective method that can be used to study gene functions in vivo,but it has some disadyantages,such as gene-trap vector fragment deletion;long fragment mising of host chromosome and wild-type transcription expression.Therefore,we must analyze the genetic background of Atpl lc gene-trap mice before studying Atpl lc biological function in vivo.In this study, the physiological state of Atpllc gene-trap mice were studied and combined with growth and development. Based on PCR, the gene-trap vector was confirmed in the position of first intron of Atp11c gene. The DNA sequencing results showed an irregular fragment deletion on the host chromosome and both ends missing of the vector.Fluorescent competitive PCR confirmed the gene-trap vector was a single copy. However Atpllc gene-trap mice have no significant difference from wild type129mice in terms of weight gaining and sexual development. We used semi-quantitative RT-PCR(reverse transcription and polymerase chain reaction) to detect Atp11c expression patterns and found the polyA sequences did not work. Atp11c gene-trap mice express both wild and mutant wild transcripts so that the phenotype of Atp11c gene-trap mice are the same as wild type mice. This may be because trapping vector insertion causes chromosome damage which results in alternative splicing, or because that the Atpl lc function is too important to the body to appear compensation splicing that ensure Atp11c expression in vivo.At the genomic level,this research have verified the gene-trap vector inserted in the first intron of Atp11c gene.The vector insertion sites and the deletion state of host chromosome have been detected.At the same time the transcripts are found diverse bands mixed together.It is confirmed that the gene-trap mice do not reach the expected purpose in Atp11c function and gene trapping has the uncertainty.