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Fosmids and BACS are cloning vectors that carry large inserts Size of partially digested genomic DNA Сopy number Number of clones for 1x human coverage Vector Large-insert clone Amp Cos site COS site Human - insert comprises Fosmid Poly- 35-45 kb Single сору -80% of -75,000 linker fosmid F-factor Cam origin of replication Cam" Human -insert Poly- 100-200 kb linker comprises -90% of BAC Single сорy 15,000- 30,000 BAC F-factor origin of replication Amp FIGURE 10-10 Features of some large-insert cloning vectors. The number of clones needed to cover the human genome once (1x) is based on a genome size of 3000 Mb (3 billion base pairs). introduce these big pieces of recombinant DNA into recipient E. coli cells. After they are in the cell, these hybrids, just like the à phage, form circular molecules that replicate extrachromosomally in a manner similar to plasmids. However, because of the presence of F plasmid origins of replications that couple plasmid replication to host cell chromosome duplication, very few copies of fosmids accu- mulate in a cell. The most popular vector for cloning very large DNA inserts in bacteria is the bacterial artificial chromosome (BAC). Derived from the F plasmid, it can carry inserts ranging from 100 to 200 kb, although the vector itself is only -7 kb (see Figure 10-10). The DNA to be cloned is inserted into the plasmid, and this large circular recombinant DNA is introduced into the bacterium. BACS were the "workhorse" vectors for the extensive cloning required by large-scale genome- sequencing projects, including the public project to sequence the human genome (discussed in Chapter 14). 00

Fosmids and BACS are cloning vectors that carry large inserts Size of partially digested genomic DNA Сopy number Number of clones for 1x human coverage Vector Large-insert clone Amp Cos site COS site Human - insert comprises Fosmid Poly- 35-45 kb Single сору -80% of -75,000 linker fosmid F-factor Cam origin of replication Cam" Human -insert Poly- 100-200 kb linker comprises -90% of BAC Single сорy 15,000- 30,000 BAC F-factor origin of replication Amp FIGURE 10-10 Features of some large-insert cloning vectors. The number of clones needed to cover the human genome once (1x) is based on a genome size of 3000 Mb (3 billion base pairs). introduce these big pieces of recombinant DNA into recipient E. coli cells. After they are in the cell, these hybrids, just like the à phage, form circular molecules that replicate extrachromosomally in a manner similar to plasmids. However, because of the presence of F plasmid origins of replications that couple plasmid replication to host cell chromosome duplication, very few copies of fosmids accu- mulate in a cell. The most popular vector for cloning very large DNA inserts in bacteria is the bacterial artificial chromosome (BAC). Derived from the F plasmid, it can carry inserts ranging from 100 to 200 kb, although the vector itself is only -7 kb (see Figure 10-10). The DNA to be cloned is inserted into the plasmid, and this large circular recombinant DNA is introduced into the bacterium. BACS were the "workhorse" vectors for the extensive cloning required by large-scale genome- sequencing projects, including the public project to sequence the human genome (discussed in Chapter 14). 00

Biology 2e
Biology 2e
2nd Edition
ISBN:9781947172517
Author:Matthew Douglas, Jung Choi, Mary Ann Clark
Publisher:Matthew Douglas, Jung Choi, Mary Ann Clark
Chapter14: Dna Structure And Function
Section: Chapter Questions
Problem 18RQ: During proofreading, which of the following enzymes reads the DNA? primase topoisomerase DNA pol...
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In Figure 10-10, determine approximately how many
BAC clones are needed to provide 1× coverage of
a. the yeast genome (12 Mbp).
b. the E. coli genome (4.1 Mbp).
c. the fruit-fly genome (130 Mbp).

Fosmids and BACS are cloning vectors that carry large inserts Size of partially digested genomic DNA Сopy number Number of clones for 1x human coverage Vector Large-insert clone Amp Cos site COS site Human - insert comprises Fosmid Poly- 35-45 kb Single сору -80% of -75,000 linker fosmid F-factor Cam origin of replication Cam" Human -insert Poly- 100-200 kb linker comprises -90% of BAC Single сорy 15,000- 30,000 BAC F-factor origin of replication Amp FIGURE 10-10 Features of some large-insert cloning vectors. The number of clones needed to cover the human genome once (1x) is based on a genome size of 3000 Mb (3 billion base pairs). introduce these big pieces of recombinant DNA into recipient E. coli cells. After they are in the cell, these hybrids, just like the à phage, form circular molecules that replicate extrachromosomally in a manner similar to plasmids. However, because of the presence of F plasmid origins of replications that couple plasmid replication to host cell chromosome duplication, very few copies of fosmids accu- mulate in a cell. The most popular vector for cloning very large DNA inserts in bacteria is the bacterial artificial chromosome (BAC). Derived from the F plasmid, it can carry inserts ranging from 100 to 200 kb, although the vector itself is only -7 kb (see Figure 10-10). The DNA to be cloned is inserted into the plasmid, and this large circular recombinant DNA is introduced into the bacterium. BACS were the "workhorse" vectors for the extensive cloning required by large-scale genome- sequencing projects, including the public project to sequence the human genome (discussed in Chapter 14). 00
Transcribed Image Text:Fosmids and BACS are cloning vectors that carry large inserts Size of partially digested genomic DNA Сopy number Number of clones for 1x human coverage Vector Large-insert clone Amp Cos site COS site Human - insert comprises Fosmid Poly- 35-45 kb Single сору -80% of -75,000 linker fosmid F-factor Cam origin of replication Cam" Human -insert Poly- 100-200 kb linker comprises -90% of BAC Single сорy 15,000- 30,000 BAC F-factor origin of replication Amp FIGURE 10-10 Features of some large-insert cloning vectors. The number of clones needed to cover the human genome once (1x) is based on a genome size of 3000 Mb (3 billion base pairs). introduce these big pieces of recombinant DNA into recipient E. coli cells. After they are in the cell, these hybrids, just like the à phage, form circular molecules that replicate extrachromosomally in a manner similar to plasmids. However, because of the presence of F plasmid origins of replications that couple plasmid replication to host cell chromosome duplication, very few copies of fosmids accu- mulate in a cell. The most popular vector for cloning very large DNA inserts in bacteria is the bacterial artificial chromosome (BAC). Derived from the F plasmid, it can carry inserts ranging from 100 to 200 kb, although the vector itself is only -7 kb (see Figure 10-10). The DNA to be cloned is inserted into the plasmid, and this large circular recombinant DNA is introduced into the bacterium. BACS were the "workhorse" vectors for the extensive cloning required by large-scale genome- sequencing projects, including the public project to sequence the human genome (discussed in Chapter 14). 00
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