Chen Lit Yang

A133282

GENE EXPRESSION TUTORIAL

1.

Compare the differences in regulation of gene expression between prokaryotes and eukaryotes. (10m) Regulations of gene expressed in prokaryote most likely occur during the process of transcription due to the need of prokaryotes to respond quickly to their environment when changes occur. Translational control also occurs, mediated by stability of the mRNAs. Regulations of prokaryotes in gene expression are shown by the operon model where it is self-regulating series of genes that work together. An operon includes a special segment of genes that are regulators of the protein synthesis, but do not code for protein, called the promoter and operator. These segments overlap and their interaction determine whether the process will start and when it will stop. Besides, negative Prokaryotic gene regulation will occur and usually comes in the form of feedback inhibition, with the end product of the process either interfering with an enzyme early in the production process, or by directly interfering with the operator sequence, acting as a repressor. There is also positive gene regulation found in prokaryotes. In this case, a repressor is normally attached to the operator in nucleotide sequence. A protein or a smaller molecule will act as an activator, attaching to the repressor and de-activating it. Unlike prokaryotes, regulation of eukaryotes in gene expression can be controlled at various location and have multiple gene-regulating mechanism that operate in the nucleus before and after RNA transcription, and in the cytoplasm both before and after translation. This is because eukaryotes have compartments such as nucleus within cells, and often multicellular structures that require differentiation of cells. Regulation of gene expression in eukaryotes occur in several main levels include genome, transcription, processing of RNA, translation of protein, posttranslational events and degradation of mRNA and proteins. Eukaryotic genes have a TATA box right before and enhancers very far upstream from the promoter sequence. When activators attach to the enhancer regions, a group of mediator proteins and some transcription factors will fall into place, allowing RNA polymerase to attach. This forms the transcription initiation complex. Without any of these elements, the gene cannot be transduced. In this way, eukaryotic genes may be regulated.

2.

Predict and briefly explain the effect on GAL1 transcription (increase/decrease) in the galactose alone, of the following mutation.

in

yeast

a) Addition of six Gal4 binding sites to the Gal 1 UAS element. (2m) GAL 1 transcription in yeast will increase with addition of six Gal4 binding sites to the Gal1 UAS element in the presence of galactose. This is because Galp acts as a transcription activator and with presence of galactose, transcription is turn on. b) Mutation occur at the gene encodes for Gal 80p. (2m) Gal 1 transcription in yeast will increase. If mutations occur at gene encoding Gal80p means that Gal80p will not be produce. Gal80p act as a repressor which binds to Gal4p

which will block the activation domain of the Gal4p. Without Gal80p, transcription is turn on and activated by Gal4p.

3.

Predict and briefly explain the effect on Lac Z transcription in E.coli (increase/decrease), in the presence of lactose alone, of the following mutation: a) Deletion of the Lac I gene (2m) The regulatory gene is the I gene that encodes for repressor protein of the Lac operon. Hence, deletion of the LacI gene will increase LacZ transcription in E.coli. b) Mutation occur at the Lac A gene (2m) There will not be any effect on lac Z transcription in E.coli if mutation occurs at the lac A gene. This is because lac A gene is located downstream from lac Z gene and the lac Z gene will still be transcribe by RNA polymerase. c) Nucleotide sequence changes at the operon site. (2m) The Lac Z transcription in E.coli will increase with nucleotide sequence changes at the operon site. This is because with mutation at the operator site, the lac repressor cannot bind to the operator and the transcription will occur.