y****e
发帖数: 71 | 1
Why not? Use reverse transcriptase just few days ago, to make cDNA from
mRNA.
A lot of RNA viruses use RT when infect host, and integrate into host genomic
DNA. | y****e
发帖数: 71 | | a****a
发帖数: 3905 | 3 sequence the protein, then make the cDNA, although u
may loss some part of the DNA. | y****e
发帖数: 71 | 4
Protein sequence is very hard to do. In fact, people all sequence DNA, then
deduce protein sequence.
Also, you might form unstable mRNA, since most amino acids have more than
one
codon, you do not know which one you should use.
Even though, people still did this in small peptides synthesis. However, I
did not hear anyone do the same thing to big proteins.
【在 a****a 的大作中提到】
: sequence the protein, then make the cDNA, although u
: may loss some part of the DNA.
| k***r
发帖数: 100 | 5 The techon. now can synthesis up to 300 A.A. peptide. with >99% purity.
But very very very expensive!
【在 y****e 的大作中提到】
:
: Protein sequence is very hard to do. In fact, people all sequence DNA, then
: deduce protein sequence.
: Also, you might form unstable mRNA, since most amino acids have more than
: one
: codon, you do not know which one you should use.
: Even though, people still did this in small peptides synthesis. However, I
: did not hear anyone do the same thing to big proteins.
| y****e
发帖数: 71 | 6
You are misunstanding here. We are talking about knowing peptide sequence,
reverse engeneering DNA sequence which can make this peptide in vivo.
Chemical synthesis protein is a hell of work.
【在 k***r 的大作中提到】
: The techon. now can synthesis up to 300 A.A. peptide. with >99% purity.
: But very very very expensive!
| s*k
发帖数: 144 | 7
You mean reverse translation?--difficult in vivo.
translation is one of the most complex processes in cell. | a****a
发帖数: 3905 | 8 You neednot to seq. all peptide la. Seq. some fragment and try to fish out
from lib. Anyway, there are a lot of things may affect the result, but it is
still a possible way.
【在 y****e 的大作中提到】
:
: You are misunstanding here. We are talking about knowing peptide sequence,
: reverse engeneering DNA sequence which can make this peptide in vivo.
: Chemical synthesis protein is a hell of work.
| y***n
发帖数: 99 | 9 The routin is to sequence C-terminal ~7aa. Then find out the corresponding
DNA sequence. Besides searching the genome sequenced, you can do reverse
PCR using wobbling codon to get the corresponding cDNA from total mRNA. Then,
clone it and do the expression.
【在 a****a 的大作中提到】
: You neednot to seq. all peptide la. Seq. some fragment and try to fish out
: from lib. Anyway, there are a lot of things may affect the result, but it is
: still a possible way.
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