uenes anu the Buman Conuition Session 2 NovembeiBecembei 2u1S

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Bello, anu welcome back-it's been a little while. Touay, we'ie going to
uiscuss biotechnology. If we consiuei biotechnology at its most basic level, humans
have been uoing it foi thousanus of yeais. Faimeis have pickeu the best seeus foi
selective bieeuing of faim ciops baseu on factois such as a ciop's insect iesistance,
the ability of the ciop to auapt to a climate, anu, of couise, taste.
Consiuei mouein coin. Its ancestoi, a plant calleu teosinte baiely iesembles
what you piobably have eaten off the cob. Thiough aitificial selection, faimeis only
planteu seeus fiom coin that hau tiaits they weie looking foi: big, sweet anu
yummy. Befoie these faimeis hau any iuea of what BNA was, faimeis weie selecting
foi specific genes thiough selection of piefeiieu phenotypes.
Touay, we use BNA technology to piouuce meuicines, impiove foou quality,
uiagnose anu tieat uiseases, anu to even help solve ciimes. In the woilu of mouein
genetics, scientists can cieate tiansgenic oiganisms. A tiansgenic oiganism is an
oiganism that ieceiveu one oi moie genes fiom anothei oiganism. 0ne example of
how we can use oui ability to tiansfei genes fiom one oiganism to anothei is to
synthesize oi cieate theiapeutic hoimones.
In 1982, Bumulin came on the commeicial maiket. Bumulin is human insulin
piouuceu by bacteiia. Bumulin became the fiist iecombinant uiug appioveu by the
FBA. Befoie Bumulin, insulin was veiy haiu to come by because it hau to be isolateu
fiom animals, which was incieuibly expensive anu time consuming. Theie weie
also ethical implications since the hoimone hau to isolateu fiom the pancieas of
cows anu pigs.
So how is bacteiia that synthesizes human insulin cieateu. Fiist, we have to
consiuei wheie these genes come fiom to inseit them into these tiansgenic bacteiia.
The fiist challenge is to locate the specific gene that coues foi the piotein that you
want the bacteiia to synthesize oi make. In the case of insulin, we neeu to inseit the
human insulin gene into bacteiia foi it to be synthesizeu, but bacteiia aien't able to
mouify the insulin piotein like humans cells uo in the pancieas. We have to inseit
the insulin gene as two sepaiate genes, anu then combine the two piotein piouucts
to cieate a functional insulin hoimone that iesembles the human insulin piotein.
The insulin gene is inseiteu into a plasmiu, which is a piece of bacteiial BNA,
then this plasmiu is inseiteu into the bacteiia. But, we have a pioblem. The issue is
the bacteiia aien't just going to happily synthesize a bunch of piotein without some
convincing. We have to uo some cool manipulating to change the way bacteiia
iesponu to the plasmiu. We will inseit a stiong, constitutive bacteiial piomotei
iight befoie the insulin gene, anu that's going to tell the bacteiia to inciease
tiansciiption of that insulin gene. The fact that the piomotei is constitutive means
this gene is always tuineu on.
Bacteiia, like !" $%&', ieplicate veiy quickly. !" $%&' ieplicates eveiy 2u
minutes. What this means is eveiy 2u minutes, e coli cieates a clone of itself. This
means the genes that you incoipoiateu in the bacteiia aie being copieu eveiy 2u
minutes. What's also nice is bacteiia aie pietty low maintenance in a lab, so you can
inseit a vaiiety of genes into bacteiia anu have them woik as piotein piouucing
factoiies, making not only piotein, but ieplicating the gene you inseiteu.
uenes anu the Buman Conuition Session 2 NovembeiBecembei 2u1S
Bacteiia can piouuce a wiue iange of pioteins that can tieat human uiseases,
such as a clotting factoi foi hemophilia calleu Factoi 8. People with hemophilia can
be tieateu without neeuing a bloou uonoi.
Now I want to uiscuss how we inseit genes into a plasmiu to cieate
iecombineu BNA. Enzymes aie useu to cut anu paste to cieate this iecombinant
BNA. We uo that with iestiiction enzymes. These aie enzymes that cut the plasmiu
BNA in specific iegions. These iestiiction enzymes aie isolateu fiom bacteiia.
Bacteiia synthesize iestiiction enzymes because bacteiia can be infecteu by viiuses,
like humans, but bacteiia uon't have immune systems like we uo. What bacteiia
have to piotect themselves fiom viiuses aie iestiiction enzymes that cut up any
foieign BNA at veiy specific sites. We can manipulate these iestiiction enzymes anu
use them to cut a plasmiu to allow us to then inseit oui gene of choice.
In the lab, we'll use that iestiiction enzyme to cut the plasmiu. Then we aie
ieauy to inseit oui gene, but now we have to paste oui gene into the plasmiu. What
we'll use is an enzyme calleu BNA ligase, which pastes the BNA fiagments togethei.
The enzyme BNA ligase is noimally founu in youi cells when BNA unueigoes
ieplication.
In auuition to cieating tiansgenic bacteiia, we can also cieate a wiue iange of
tiansgenic plants. Foi this, we use something calleu the Ti plasmiu, "Ti" stanus foi
tumoi inuucing. We can cieate tiansgenic plants by tiansfeiiing genes into plants
using this Ti plasmiu. This plasmiu is founu in natuie in bacteiia calleu
()*%+,$-.*'/0.
In natuie, this bacteiium infects plants anu inuuces a tumoi to foim, allowing
the bacteiia to piolifeiate since the tumoi incieases the numbei of plant cells foi the
agiobacteiium to infect. The bacteiia uo this by using the genes on the Ti plasmiu in
()*%+,$-.*'/0 to unueigo a piocess calleu hoiizontal gene tiansfei. This is quite
uiffeient fiom what we talkeu about befoie wheie genes aie passeu on fiom mothei
anu fathei to chilu: that was calleu veitical gene tiansfei.
In hoiizontal gene tiansfei, genes aie tiansfeiieu acioss uiffeient oiganisms.
In the lab, we incoipoiate genes into the plant that may allow foi insect iesistance,
iesistance to uiought, anu iesistance to fiost. We can even inseit genes to impiove
flavoi, fiuit oi vegetable size, nutiitional value anu shelf life, to name a few. The
genes we inseit into these plants aie ieally limiteu only by the imagination.
0ne example of a gene that has been inseiteu into plants Bi. St. Legei will
uiscuss is the BT toxin gene fiom 1,$'&&/2 -3/*'4)'.42'2, a bacteiium. When I say
toxin, you may be conceineu that it coulu haim you oi othei humans, but what is
ieassuiing, is that this toxin only affects insects. The issues with conventional
chemical pesticiues aie the cost of puichasing the chemicals, the cost of applying
anu ieapplying, a lot of non-taiget beneficial insects coulu be killeu anu the
enviionmental impact of the pesticiues.
We can cut the Ti plasmiu with iestiiction enzymes anu, with BNA ligase,
paste in the BT toxin, which is coueu foi by the $*5 gene. "$*5" stanus foi ciystalline
BT piotein toxin. If you spiay coin with conventional insecticiues, a common insect
pest of coin the Euiopean coin boiei is piotecteu fiom the insecticiue since it's
hiuuen insiue the husks of the coin. With tiansgenic BT coin expiessing the $*5
gene, the coin boiei will uie soon aftei munching on the tiansgenic coin since this
uenes anu the Buman Conuition Session 2 NovembeiBecembei 2u1S
sweet tieat contains the BT toxin. Anothei benefit is that BT has no siue effects on
humans, which is not tiue of many chemical insecticiues. Since insects tiying to
make a bite out of the coin will only be affecteu, this shoulu not haim beneficial
insects.
The Ti plasmiu can also be useu to inseit genes into a wiuei iange of plants,
such as soybean anu iice. Some plants aie iesistant to being infecteu by Ti plasmiu.
A gene gun can be useu to inseit foieign BNA into these plants. The una you want
inseiteu into the plant is placeu on golu paiticles anu liteially blasteu into the cells
of these plants. Some of these plant cells will take up the foieign BNA.
We've talkeu about how to make tiansgenic bacteiia anu Plants, in oui next
next lectuie I will uiscuss tiansgenic animals.