18. Biotechnology

Introduction to DNA Cloning

[LECT C8 : RECOMBINANT DNA TECHNOLOGY] 8.2 Steps in Gene Cloning

 inducing recombination in differentiated donor cells in order to restore the full genomic complement 

 No. Differentiated cells already have a full genomic complement. 

 transforming donor cells with genes encoding proteins required for normal embryonic development 

 No. Currently, at least, cloning does not involve the introduction of additional genes into the donor cell nuclei. 

 physically removing the nucleus from the egg cell that will ultimately receive the donor cell nucleus 

 No. Though not necessarily straightforward, this has not been a particular problem in cloning. 

 implanting the clone into the surrogate mother 

 completely reversing epigenetic alterations in donor cell nuclei such as DNA methylation and chromatin packing 

What must occur for a differentiated cell to support embryonic development?

The efficiency of cloning, and the ability to generate healthy cloned animals, has been largely hampered by the difficulty of __________.

Which of the following is not true of cDNA produced using human brain tissue as the starting material?
a. It can be amplified by the polymerase chain reaction.
b. It was produced from pre-mRNA using reverse transcriptase.
c. It can be labeled and used as a probe to detect genes expressed in the brain.
d. It lacks the introns of the pre-mRNA.

In DNA technology, the term vector can refer to
a. the enzyme that cuts DNA into restriction fragments.
b. the sticky end of a DNA fragment.
c. a SNP marker.
d. a plasmid used to transfer DNA into a living cell.

DNA cloning

DNA cloning and recombinant DNA | Biomolecules | MCAT | Khan Academy

Which of the following is true of a codon? 
(A) It never codes for the same amino acid as another codon. 
(B) It can code for more than one amino acid. 
(C) It can be either in DNA or in RNA 
(D) It is the basic unit of protein structure

The enzyme that cuts DNA into restriction fragments.

A vehicle used to transfer DNA into a living cell.

In DNA technology, the term “vector” can refer to:

Which of the following is true of cDNA produced using human brain tissue as the starting material? 
(A) The procedure to make it requires amplification by the polymerase chain reaction. 
(B) It is produced from pre-mRNA using reverse transcriptase. 
(C) It can be labeled and used as a probe to detect genes expressed in the brain. 
(D) It includes the introns of the pre-mRNA.

Small accessory rings of DNA that replicate independently of an organism’s genome are called _____.

Animation: Recombinant DNA

 locate specific genes on animal chromosomes 

 No. Plasmids are not nucleic acid probes. 

 detect and correct mistakes in DNA replication 

 All of the listed responses are correct. 

 No. Only one of the other choices is correct. 

 insert genes of interest into plant chromosomes 

This bacterium is the source of the Ti plasmid.

In genetic engineering, the highly active plasmid from Agrobacterium tumefaciens is used to __________.

Introduction to Molecular Cloning

Recombinant DNA Process

 The virus caused the mouse cells to become diseased. 

 No. Hopefully, disease-causing genes of the virus had been removed prior to its use as a vector. 

 The virus had transferred a gene from one mouse cell to another. 

 No. This might have happened, but it is not an explanation for the results. 

 The virus was too small to carry the entire gene. 

 No. If this were so, the cell would not have been able to express the introduced gene. 

 The enzyme acted as a nuclease enzyme, cutting up mouse DNA. 

 No. This would not have led to the observed results. 

 The virus inserted the gene encoding the human enzyme within the sequence of a normally expressed endogenous gene. 

What do retroviruses do when they enter cells?

A molecular biologist used a retroviral vector to introduce a gene coding for a certain human enzyme into mouse cells. One cell line was isolated that was able to make the human enzyme, but it had lost the ability to express an endogenous, normally expressed gene in the process. What is the best explanation for these results?

 No. These enzymes do not synthesize DNA. 

 No. These enzymes do not synthesize RNA. 

 No. These enzymes do not synthesize protein. 

 No. These enzymes do not break peptide bonds. 

Consider how these enzymes are used in DNA technology.

Bacteria use restriction enzymes to __________.

Cloning with Recombinant DNA

Animation 27.1 Basic principle of recombinant DNA technology

 naturally occurring DNA molecules are very long and contain many genes 

 No. This is true, but there is a better answer. 

 it provides a means to produce large quantities of its protein product 

 genes occupy only a small proportion of the chromosomal DNA in eukaryotes, the rest being noncoding nucleotide sequences 

 it provides a means to produce many copies of a gene in a short period of time 

Consider the comparative size and frequency of a specific gene to the entire genome of an organism.

Gene cloning is crucial to any application involving one gene because __________.

A bacterial chromosome and a bacterial plasmid.

 No. Reverse transcriptase catalyzes the formation of cDNA. 

 No. Restriction enzymes cut DNA at specific recognition sites. 

 No. DNA polymerase catalyzes DNA replication. 

Review recombinant DNA technology.

Which of the following enzymes could seal a nick in one strand of a double-stranded DNA molecule by creating a sugar-phosphate bond between the adjacent, unjoined nucleotides?

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