From Gene to Protein: Translation Into Biotechnology
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- The Genetics of the Dog.
- The centrality of RNA for engineering gene expression;
- Alternative RNA Splicing.
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How Genes Are Regulated – Concepts of Biology – 1st Canadian Edition
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Request a Quote. This makes it impossible for DNA polymerases to synthesize both strands simultaneously. A portion of the double helix must first unwind, and this is mediated by helicase enzymes. The leading strand is synthesized continuously but the opposite strand is copied in short bursts of about bases, as the lagging strand template becomes available. The resulting short strands are called Okazaki fragments after their discoverers, Reiji and Tsuneko Okazaki.
Does the ‘Central Dogma’ always apply?
Pol III can then take over, but it eventually encounters one of the previously synthesized short RNA fragments in its path. The initiation of DNA replication at the leading strand is more complex and is discussed in detail in more specialized texts. DNA replication is not perfect. This leads to mismatched base pairs, or mispairs. DNA polymerases have proofreading activity, and a DNA repair enzymes have evolved to correct these mistakes.
Occasionally, mispairs survive and are incorporated into the genome in the next round of replication.
9.5 How Genes Are Regulated
These mutations may have no consequence, they may result in the death of the organism, they may result in a genetic disease or cancer; or they may give the organism a competitive advantage over its neighbours, which leads to evolution by natural selection. Transcription is the process by which DNA is copied transcribed to mRNA, which carries the information needed for protein synthesis. Transcription takes place in two broad steps.
The mechanism of transcription has parallels in that of DNA replication. As with DNA replication, partial unwinding of the double helix must occur before transcription can take place, and it is the RNA polymerase enzymes that catalyze this process. Unlike DNA replication, in which both strands are copied, only one strand is transcribed. The strand that contains the gene is called the sense strand, while the complementary strand is the antisense strand. The mRNA produced in transcription is a copy of the sense strand, but it is the antisense strand that is transcribed.
Transcription ends when the RNA polymerase enzyme reaches a triplet of bases that is read as a "stop" signal. The DNA molecule re-winds to re-form the double helix. The pre-messenger RNA thus formed contains introns which are not required for protein synthesis. In alternative splicing, individual exons are either spliced or included, giving rise to several different possible mRNA products.
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Each mRNA product codes for a different protein isoform; these protein isoforms differ in their peptide sequence and therefore their biological activity. Several different mechanisms of alternative splicing are known, two of which are illustrated in Figure 6. Splicing is important in genetic regulation alteration of the splicing pattern in response to cellular conditions changes protein expression. Perhaps not surprisingly, abnormal splicing patterns can lead to disease states including cancer.
This process, catalyzed by reverse transcriptase enzymes, allows retroviruses, including the human immunodeficiency virus HIV , to use RNA as their genetic material. The mRNA formed in transcription is transported out of the nucleus, into the cytoplasm, to the ribosome the cell's protein synthesis factory.
Here, it directs protein synthesis. The ribosome is a very large complex of RNA and protein molecules. Each three-base stretch of mRNA triplet is known as a codon , and one codon contains the information for a specific amino acid. The tRNA is then expelled from the ribosome. Figure 7 shows the steps involved in protein synthesis. Figure 7 Translation a and b tRNA molecules bind to the two binding sites of the ribosome, and by hydrogen bonding to the mRNA; c a peptide bond forms between the two amino acids to make a dipeptide, while the tRNA molecule is left uncharged; d the uncharged tRNA molecule leaves the ribosome, while the ribosome moves one codon to the right the dipeptide is translocated from one binding site to the other ; e another tRNA molecule binds; f a peptide bond forms between the two amino acids to make a tripeptide; g the uncharged tRNA molecule leaves the ribosome.
Transfer RNA adopts a well defined tertiary structure which is normally represented in two dimensions as a cloverleaf shape, as in Figure 7. The structure of tRNA is shown in more detail in Figure 8. The reaction of esters with amines is generally favourable but the rate of reaction is increased greatly in the ribosome.