CONSTRUCTION AND ANALYSIS OF ARRAYED CDNA LIBRARIES

摘要: For any attempt to understand the biology of an organism incorporation a cDNA-based approach is unavoidable, because it major studying gene function. The complete sequence genome alone not sufficient organism; its regulation, expression, splice variation, posttranslational modifications, and proteinprotein interactions all need be addressed. Because majority vertebrate genes have probably been identified as ESTs44,52 next stage Human Genome Project attributing functional information these sequences. In most cases hybridization-based approaches on arrayed pieces DNA represent efficient way study expression level splicing in given tissue. Similar technology, now being applied at protein using libraries, high-density membranes, antibody screening, should allow studies localization modifications. Coupled use technologies, which although lacking highly parallel nature hybridization, can potentially characterize large numbers samples individually with high accuracy. Automated gel-based sequencing example such technique; mass fingerprinting are further examples. case spectroscopic analysis, speed sensitivity vastly superior that approaches; however, preparation more tedious. Our laboratory developing system by spectrometry, allowing rapid genotyping than currently possible technologies (■ Gut, ■ Berlin H. Lehrach, personal communication 1998). Such technology would make polymorphisms widely accessible. Data generated techniques will connected both libraries database comparisons; finally, two hybrid library screens identify many protein-protein interactions, linking together.66–68 this we start interplay between global scale, molecular interaction biological processes they regulate.