Tecnologia de DNA Recombinante
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Questions and Answers

O vírus da dengue é classificado como um vírus de RNA +ssRNA.

True

Vírus de RNA -ssRNA, como o da raiva, não precisam ser transcritos antes da tradução de suas proteínas.

False

A transcrição em procariotos termina por uma sequência específica que forma uma estrutura de laço.

True

O evento de trans-splicing é comum em todos os eucariotos multicelulares.

<p>False</p> Signup and view all the answers

A estrutura canônica de um operon bacteriano consiste em um único promotor e múltiplos genes estruturais.

<p>True</p> Signup and view all the answers

A conclusão do Genoma Humano não permitiu identificar a presença de muitas sequências regulatórias no nosso DNA.

<p>False</p> Signup and view all the answers

O início da tradução em procariotos ocorre na presença de um códon de parada.

<p>False</p> Signup and view all the answers

A eletroforese bidimensional é uma técnica utilizada apenas para análise de ácidos nucleicos, não proteínas.

<p>False</p> Signup and view all the answers

O Projeto Genoma Humano foi finalizado e publicado em 1995.

<p>False</p> Signup and view all the answers

A técnica de 'shotgun' utilizada no sequenciamento do genoma humano é baseada em cortes aleatórios do DNA.

<p>True</p> Signup and view all the answers

O teste de PCR é uma técnica que permite a amplificação do material genético do SARS-CoV-2 antes de sua detecção.

<p>True</p> Signup and view all the answers

A insulina recombinante produzida em E. coli não precisa ser tratada antes do uso.

<p>False</p> Signup and view all the answers

A coleta de amostras para pesquisa proteômica deve ser sempre realizada após o início do tratamento oncológico.

<p>False</p> Signup and view all the answers

O operon lac é ativado em meio com glicose e lactose, permitindo a produção de lactase.

<p>False</p> Signup and view all the answers

As enzimas polimerases são responsáveis pela degradação do material genético do SARS-CoV-2 durante a PCR.

<p>False</p> Signup and view all the answers

A transcrição reversa é uma etapa que não pode ser utilizada na produção de proteínas recombinantes em eucariotos.

<p>False</p> Signup and view all the answers

Os pseudogenes são segmentos do DNA que possuem funções importantes conhecidas.

<p>False</p> Signup and view all the answers

A resistência à ampicilina em Pseudomonas aeruginosa está relacionada à expressão do gene ampR.

<p>True</p> Signup and view all the answers

As regiões do DNA altamente repetitivas são irrelevantes para a genética humana.

<p>False</p> Signup and view all the answers

O vetor de expressão em técnicas de DNA recombinante deve conter um sítio promotor fraco.

<p>False</p> Signup and view all the answers

Study Notes

DNA Recombinant Technology

  • DNA recombinant technology allows joining different DNA molecules using restriction enzymes and DNA ligase.
  • Different restriction sites allow unidirectional insertion of the DNA of interest.
  • Recombinant DNA is transferred to a microtube for in vitro protein transcription and translation.
  • The protein of interest, after large-scale production, purification, and denaturation, can be used as a vaccine antigen, hormone, or recombinant enzyme.

Hearing Loss Diagnosis

  • A 7-week-old girl (A.J.S.) was diagnosed with congenital (non-syndromic) hearing loss.
  • Her parents had no history of hearing problems.
  • Hearing tests revealed bilateral hearing loss.
  • A molecular analysis identified homozygous mutations in the autosomal gene GJB2.
  • GJB2 codes for connexin 26, a protein responsible for gap junctions in cochlear cells.
  • The GJB2 gene sequence was analyzed comparing the wild-type allele with the mutated allele.

Molecular Techniques for Diagnosis

  • PCR followed by restriction and electrophoresis of the GJB2 gene can be used for molecular diagnosis.
  • DNA extraction from the patient is a necessary step before applying any molecular test.

Human Genome Sequencing

  • Human genome sequencing used restriction enzymes and shotgun sequencing-based techniques.
  • The project identified pseudogenes, highly repetitive DNA regions, and numerous regulatory sequences.
  • Gene families are organized in operons.
  • This study was significant for medical genetics, contributing to a better understanding of diseases and potential treatments.

Pseudomonas aeruginosa Resistance Analysis

  • Miguel isolated 17 ampicillin-resistant Pseudomonas aeruginosa strains.
  • He used plant extract from cashew tree bark to evaluate its effect on the ampR gene expression.
  • PCR is a useful technique to study gene expression.

Operon Lac Regulation

  • When a bacterium is cultivated in a medium containing glucose and lactose, the lac operon expression decreases.

DNA Recombinant Technology Assessment

  • Generating a recombinant virus protein using E. coli as a host cell is possible.
  • Different restriction sites are needed for each gene of interest.
  • The expression vector is a plasmid with a strong promoter.
  • Reverse transcription can be used in the production of eukaryotic recombinant proteins.
  • Recombinant insulin does not need antibiotic pre-treatment.

DNA Fragment Analysis in a Crime Scene

  • Fragments of a 3-year-old girl were found at a crime scene.
  • DNA analysis of the mother and fragments were compared, confirming that the father is not the child's biological father.
  • Fragments 01, 04, and 05 belonged to the girl. This eliminates the other possibilities.

Viral Infection Mechanisms

  • Differences in infection and replication mechanisms exist between +ssRNA (e.g. dengue) and -ssRNA (e.g. rabies) viruses.

Transcription Termination in Prokaryotes

  • Transcription termination in prokaryotes involves Rho factor detaching the RNA polymerase from the DNA strand.

Splicing in Unicellular Eukaryotes

  • Trans-splicing is an event where different RNA molecules are spliced together, present in some unicellular eukaryotes.

Bacterial Operon Structure

  • A bacterial operon has a promoter, operator, structural genes, and a regulator.

Protein Translation Initiation in Prokaryotes

  • Translation initiation in prokaryotes involves the ribosome binding to the Shine-Dalgarno sequence (RBS).

Osteosarcoma Proteomic Analysis

  • A 24-year-old male patient has a maxillar osteosarcoma.
  • 2D electrophoresis was used for proteomic analysis of the tumor and healthy tissue.
  • The procedure involves protein separation based on isoelectric point and molecular weight.

COVID-19 Genetic Diagnosis

  • PCR is a genetic test for COVID-19 diagnosis, using a polymerase enzyme to detect SARS-CoV-2 RNA.
  • RNA samples are necessary.
  • A crucial step prior to PCR is the isolation of viral RNA.

HIV-1 Viral Load Quantification

  • qPCR (quantitative PCR in real-time) can be used to quantify the free viral load in HIV-1 patients.

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Description

Explore os fundamentos da tecnologia de DNA recombinante, incluindo a união de moléculas de DNA utilizando enzimas de restrição e ligase. Aprenda sobre a inserção de DNA de interesse e sua aplicação na produção de proteínas, vacinas e hormônios.

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