Human Gene Structure 3.pdf
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San Lorenzo Ruiz College of Ormoc, Inc.
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Summary
This document provides an overview of human gene structure, transcription, and translation. It explains the process and components involved in these biological phenomena, covering the central dogma of molecular biology. These concepts are essential to understanding fundamental biological processes.
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211 Transcription REMEMBER! Nu J.eotide Bases Tranacrlpdon R"IA G-CAT AAAA The Central Dogma of HUMAN GENE STRUCTURE Molecular Biology 1. Genes G a. Exons separated by introns b. Sequence of base pairs encodes NA.. RNA.. Protein information for proteins. replication transcription translation c. Different sequence= different p rotein TRANSLATION / GENETIC CODE 1. E ach combination of 3 nucleotides on.J u._ ,...,e:.,._...__, _______,_ o_e_, "~----~-tr_~_c1 - _~_r:_. _,,~ mRNA is called codon 2. Each codon specifies a particular amino Exo'I acid ~~ !!=Jr~ 3. Each amino acid is to be placed in the If/ polypeptide chain (protein) TRANSCRIPTION ~ 1. mR NA sequence - complementary to 5· NNNNNNN;;;c7AGCCA~ = 3· the DNA template a. Uracil ( U) bases replace thymine (T) bases in RNA Codon Usage AUG-CUC-GGG-AGC-CAU-UAA 2. mRNA processed by transcription: a. Splicing - r emoval of introns Translation b. Capping - modify the 5 ' end ♦ c. Polyadenylations - add adenines to Peptide S equence Met-Lcu-Glv~Ser-His the 3 'end (also called poly-A tail) @ REMEMBER! INtrons remain IN the nucleus; EXonsEXit Lpl--I ~ Telopl--I C Molecular Techniques @--· @)@) BASIC STEPS IN ISOLATING DNA FROM CLINICAL SPECIMENS 0 -pi--r Separate WBCs from RBCs, if necessary t lyse WBCs or other nucleated cells t Denature / Digest proteins t Separate contaminants (e.g., proteins, heme) from D NA t Precipitate DNA, if necessary t Resuspend DNA in final buffer DNA ISOLATION METHODS 1. Liquid phase organic extraction (phenol/chloroform) 2. Liquid phase non-organic extraction CHROMOSOMAL CROSSOVER 3. Solid phase procedures (GENETIC RECOMBINATION) ISOLATION AND PURIFICATION OF MJCLEIC ACID 1. Two chromosomes (paired up during 1. Sample source: prophase I of meiosis) exchange some a. Any tissue with nucleus portion of their DNA b. Blood collected in anticoagulant 2. Matching r egions of matching ❖ EDTA preferred chromosomes break then reconnect to ❖ DO NOT FREEZE WHOLE other chromosome (ex change of gen es) BLOOD 2. Storage conditions a. Store DNA in TE buffer at 4°C for weeks and at -20°C to - 70°C- for long term b. Store RNA in RNase free ultrapure water at - 70°C
