Gene splicing by fusion PCR is a versatile and widely used methodology, especially in synthetic biology. We here describe a rapid method for splicing two fragments by one-round fusion PCR with a dual-asymmetric primers and two-step annealing (ODT) method. During the process, the asymmetric intermediate fragments were generated in the early stage. Thereafter, they were hybridized in the subseque...

Figure 1. Diagram of a split-marker gene replacement strategy using fusion-PCR (A) FPCR step 1 amplification of flanking regions and partial marker segments. Dotted tails indicate the overlap sequences on selected primers which allow fusion in step 2. (B) FPCR step 2 flanking regions are fused to the marker segments at the overlap sights, forming two constructs. The fusion products are then amp...

Dear Editor In a recent issue of Annals of Laboratory Medicine, Shin and colleagues described two cases of Philadelphia chromosomepositive (Ph+) ALL expressing e1a3 BCR-ABL1 gene fusion product and listed other cases reported thus far that involve this fusion [1]. A basic literature review using the search term “e1a3” reveals two additional adult cases that should have been included in this lis...

PURPOSE Early detection of prostate cancer can increase the curative success rate for prostate cancer. We studied the diagnostic usefulness of TMPRSS2-ERG fusion transcripts as well as the combination of prostate cancer antigen 3 (PCA3) RNA and TMPRSS2-ERG fusion transcripts in urinary sediments after digital rectal examination (DRE). EXPERIMENTAL DESIGN A total of 78 men with prostate cancer...

We analyzed the mRNA expression of the FHIT gene by reverse transcription-PCR (RT-PCR) in 54 cases of acute lymphoblastic leukemia (ALL; 11 cases of T-cell ALL [T-ALL] and 43 cases of non-T-ALL) and 40 cases of acute myeloid leukemia (AML). In 46% of the ALL cases and 55% of the AML cases, FHIT expression was absent or markedly decreased. Only abnormal short bands were detected in 30% of the AL...

background: the purpose of this study was to clone, express, and purify a novel multidomain fusion protein of micobacterium tuberculosis (mtb) in a prokaryotic system. methods: an hspx/esxs gene construct was synthesized and ligated into a pgh plasmid, e. coli top10 cells were transformed, and the vector was purified. the vector containing the construct and pet-21b (+) plasmid were digested wit...

Background. Synovial sarcoma is an aggressive soft-tissue malignancy. This study examines the presence of the SYT-SSX fusion transcript in synovial sarcoma microvesicles as well as its potential role as a biomarker for synovial sarcoma. Patients and Methods. Microvesicle release of synovial sarcoma cells was examined by transmission electron microscopy. RNA-content was analyzed by qPCR, nested ...

BACKGROUND PML/RARalpha fusion transcripts provide a readily accessible marker for diagnosis of acute promyelocytic leukemia (APL) and for monitoring response to therapy. Survival rates are improved by therapies guided by such monitoring. We assessed the potential of DzyNA reverse transcription-PCR (RT-PCR) for measurement of PML/RARalpha fusion transcripts. METHODS Parallel single-tube DzyNA...

The characteristic t(15;17) of acute promyelocytic leukemia (APL) fuses the retinoic acid receptor alpha (RAR-alpha) gene on chromosome 17 to a gene on chromosome 15 called PML, a putative transcription factor. This distinct translocation results in a fusion mRNA detected by Northern analysis. Two cDNAs have been isolated that differ in the extent of 3' PML nucleic acid sequence contained. This...

BACKGROUND The advent of genomics-based technologies has revolutionized many fields of biological enquiry. However, chromosome walking or flanking sequence cloning is still a necessary and important procedure to determining gene structure. Such methods are used to identify T-DNA insertion sites and so are especially relevant for organisms where large T-DNA insertion libraries have been created,...