སྟོན་རྐྱངམ་ཅིག་
0:00
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Hello and welcome back to our quest to prove that we can make a vaccine for canine
0:04
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coronavirus in just 100 days.
0:06
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
This week,
0:07
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
the viral genes being used to make the vaccine arrived a whole day ahead of schedule,
0:11
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
so the team was in a flurry to get everything ready for the next steps.
0:15
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
So basically this week we received the gene box we ordered and all
0:19
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
our primers.
0:19
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
So we've been able to start our planning process.
0:23
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
So getting the inserts ready for all the different constructs
0:27
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
that were designed.
0:28
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
So we started that yesterday.
0:30
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
So how does this tiny amount of raw genes get processed not only into
0:34
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
a vaccine, but into enough vaccine for hundreds of dosages?
0:38
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
It all starts with gene cloning.
0:40
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
You may have heard of cloned animals like Dolly the Sheep or seen movies and
0:44
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
TV shows where they're cloning genes.
0:45
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
But the real process into making a vaccine is a completely different story.
0:49
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
The first step is to take these gene blocks and rehydrate them.
0:52
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
So we're cloning basically the fusion protein,
0:56
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
so the specific small part of the virus that we
1:00
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
use to...
1:01
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
pre -act our antigen,
1:03
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
so it's not the whole virus,
1:04
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
it is a subunit of it,
1:06
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
and it is basically that sequence combined with the molecular
1:10
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
clamp.
1:11
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Now,
1:11
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
here comes the cool part,
1:12
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
taking our gene construct and amplifying it.
1:15
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
If we wanted to make,
1:17
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
say, thousands of copies of a mug,
1:19
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
we would have to have a factory to make them.
1:21
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
But when we're making copies of our genes,
1:24
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
we use E.
1:25
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli as our cellular machines.
1:27
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Bacteria like E.
1:28
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli have the ability to uptake genetic material in the form of plasmids,
1:32
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
copy it for their own use,
1:33
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
and spread it to other E.
1:34
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli.
1:35
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
It's like the plasmid is a DVD with an awesome movie on it,
1:39
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
and all of the E.
1:40
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli are pirating that movie and then passing it along to their friends,
1:43
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
creating thousands and thousands of copies of our movie in the process.
1:47
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
So all we need to do is make our genetic material in the form of a
1:51
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
lot of DVDs for.
1:53
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
To make enough plasmids for our movie to go viral or maybe bacterial,
1:57
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
we need to use PCR or a polymerase chain reaction.
2:01
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
PCR is an incredibly big part of what we do.
2:04
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
So whenever we create an antigen,
2:07
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
which is the small part of the virus that we use that we put in
2:11
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
the molecular clamp.
2:13
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
Sometimes you need to do slight modifications for it to make it more stable
2:17
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
or to make sure it expresses better.
2:21
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
So we have our gene blocks,
2:23
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
which are basically almost a native sequence,
2:25
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
and then we use different primers to change small things in it.
2:29
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
And that stands for a PCR.
2:30
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Usually when cells are dividing,
2:32
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
they need to copy their DNA first with the enzyme polymerase.
2:36
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
What this reaction does is take a polymerase from a bacteria native to hot
2:41
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
springs and put it in a solution with our gene block that we need to copy.
2:44
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
We add in primers,
2:46
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
small segments of DNA that matches where we want polymerase to
2:50
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
replicate and acts as a homing beacon for the polymerase.
2:53
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Because the polymerase is from a hot spring bacteria,
2:56
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
we can warm up the DNA to split it for replicating,
2:59
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
cool it down so the primers can attach,
3:01
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
and then warm it back up again so that the bacterial polymerase can duplicate it all.
3:05
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Each cycle of this doubles the DNA,
3:07
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
so we quickly get enough DNA for all of our plasmids.
3:11
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Then, after we check the PCR copied the correct sequences,
3:13
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
we cut and glue the different sections together into a plasmid using a bunch
3:17
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
of other enzymes.
3:18
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
It's like we're editing the movie,
3:20
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
cutting and ordering the scenes to make a movie that we can put onto our DVD.
3:25
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Then we get our DVDs,
3:27
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
our plasmids,
3:27
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
and we put them in a solution with our E.
3:30
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli, heating the solution so that the plasmids are uptaken.
3:33
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Now, all that was left was for the team to let all the E.
3:35
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli grow on agar plates and incubate.
3:37
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
And every time they replicate,
3:39
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
they're copying our plasmid and sharing it.
3:42
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
Then go through the process of transformation where we put
3:46
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
the DNA into E.
3:47
S… Speaker 2 (Video 2 v2 - PCR and Cloning)
coli cells.
3:48
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
It's through this process that the team turns a tiny little pellet of
3:52
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
DNA into enough genetic material that next week...
3:54
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
they can start turning these genes into protein for the vaccine.
3:57
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
At the moment,
3:58
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
we're currently transforming those into E.
4:00
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli bacteria,
4:01
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
which we all played on agar plates overnight and purify out the DNA.
4:05
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Then we will transfect their DNA into trocells.
4:08
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
But sadly,
4:09
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
we have to wait a while for all of that E.
4:11
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli to grow.
4:11
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
So that's it for today.
4:13
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
Tune in next week to see what the team does with all of that E.
4:16
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
coli on their journey to create a vaccine in less than 100 days.
4:19
S… Speaker 1 (Video 2 v2 - PCR and Cloning)
See ya!

ཡིག་སྒྱུར་འདི་ བཅོས་མའི་བློ་རིག་ (སྒྲ་ངོས་འཛིན་བྱེད་པའི་འཕྲུལ་རིག་) གིས་བཟོ་སྟེ་ཡོདཔ་ཨིན། འཛོལ་བ་འབྱུང་སྲིད་པ་ལ་ — གལ་སྲིད་ཁག་ཆེ་བའི་དོན་ལས་ ངོ་མ་གི་སྒྲ་སྐད་དང་འཕྱད་ཞིབ་འབད་དགོཔ་ཨིན། བཅོས་མའི་བློ་རིག་གི་སྲིད་བྱུས་

❤️ STT.ai ལ་དགའ་བ་ཡིན་ནོ? སླབ་པར་བགྱི!
བཅུད་དོན་
ཡིག་ཆ་འདི་གི་ AI བཅུད་བསྡུ་ཐོན་ནི་གི་དོན་ལས་ བཅུད་བསྡུ་ ཟེར་བའི་བསྒང་འདི་བསྒང་བསྒ
དྲན་ཐོ་བསྡུ་གསོག...
དྲན་ཤེས་ལ་དྲི་བ་དྲིས་ལན་ཞུ།
ཡིག་ཆ་འདི་གི་སྐོར་ལ་དྲི་བ་ཅི་རིགས་ཞུས་ནའང་ བཅོས་མའི་བློ་རིག་གིས་ འབྲེལ་བ་ཡོད་པའི་དོན་ཚན་ཚུ་ འཚོལ་ཞིབ་ དང་ལན་འདེབས་ འབད་འོང་།