1. Two Novel Techniques Of Screening Of Gene Mutations And The Gene Diagnosis Of Chinese Hemjophilia B Patients 2. Expression Of PAL CDNA From Parsley In Lactococcus Lactis And It's Application To PKU Gene Therapy As A New Strategy

Subject I : Two Novel Techniques of Screening of Gene Mutations and the Gene Diagnosis of Chinese Hemjophilia B PatientsThe screening of gene mutations has become a major focus for the gene diagnosis and prenatal diagnosis . First we set up two rapid and sensitive methods for screening of unknown gene mutations. One is Cycle-dideoxy Fingerprinting (Cycle-ddF), a combination between the dsDNA cycle-sequencing and SSCP that can direct detect the presence of single base mutation and other sequence changes in PCR-amplified segments. Comparing to ddF, the main advantage of Cycle-ddF is that it omited the T7 RNA polymerase transcription and the AMV reverse transcription , decreased the cost of screening test and remained the high efficience. The other one is named as fluorescent fingerpriting(FF). This techniques is based on the fluoresent detection system and the data collection programs of ABI 373 DNA sequencer, The FF contains all samples' all raw sequence data acquired by the data collection program. In the FF, any colour difference between 2 or more lanes predicts bases change in sequences of the samples. As compared to automated DNA sequencing for detect mutations, FF is equally accurate and more simple and efficient because all samples' gene mutations can be simultaneously revealed at once.With these two core techniques, we set up a new gene diagnosis system for Hemophilia B. The FIX gene mutations of 13 severe Hemophilia B patients had been screened. For each affected patient, one defined mutation was found. All togeher 4 micro-deletions and 7 single base substitution were identified...