Mutation Screening In CDS Of LOH12CR1 From Colorectal Carcinoma And Preparation Of Rabbit Antibody Against Human LOH12CR1 Protein

Peutz-Jeghers syndrome (PJS) is a dominantly inherited disorder characterized by multiple gastrointestinal hamartomatous polyposis and mucocutaneous melanin pigmentation. It's also known for predisposition to benign and malignant tumors in multiple organ system. The locus responsible for PJS was mapped genetically to the LKB1(also known as STK11)which is defined as a tumor suppressor. The prtein STK11 is a 433AA serine/threonine protein kinase By using the method of GST-Pull down with mass chromatographic analysis and co-immunoprecipitation, our research team has already screened and defined a new LKB1 interacting protein- LOH12CR1 in L02 cell. LOH12CR1 located on the 12p,is the candidate gene of suppressor tumor gene and highly reserved in evolution. There is a deletion heterozygote phenomenon of the region D12S89-D12S358 where LOH12CR1 is located in hematopoietic system malignant and breast,lung, ovarian and other solid tumors, and the deletion heterozygote in hematopoietic system malignant often accompanied with a phenomenon that the expression of abundance LOH12CR1 also reduced. The CDS of LOH12CR1 in 12 pairs of cancer and adjacent tissues were amplified by PT-PCR, and the products were directly sequenced in order to analyse whether there is a mutation in CDS of LOH12CR1 in these tissues.Besides,in order to know the expression level of LOH12CR1 in different tissues, first we should obtaine effective rabbit polyclonal antibody of LOH12CR1 protein as a tool to carry on our future research. We use purified LOH12CR1 protein and Freund's adjuvant mixture as antigens in rabbits,to obtain specificity Rabbit anti-human LOH12CR1 serum, then Western blot was used to examine it.