请问怎样能construct 一个heterodimer?# Biology - 生物学
h*r
1 楼
置顶帖里这么说，可是看到其他网站又说97年之后发的护照是有这个号的，到底有没有
有啊？表上怎么填啊？谢谢。
有啊？表上怎么填啊？谢谢。
n*I
2 楼
去年买了房子,交了大半年的按揭房贷.今天收到了房贷银行的信,说是用于报税的, 听
说房贷利息是减税的. 第一次买房报税,恳求过来人指点一二?谢谢!
说房贷利息是减税的. 第一次买房报税,恳求过来人指点一二?谢谢!
b*i
3 楼
是寄I-129F申请表的时候连同寄上还是等签证中心通知收到表以后再寄？
还有各自陈述双方怎样认识的信如果对方英文不好可以用中文写吗？
谢谢！
还有各自陈述双方怎样认识的信如果对方英文不好可以用中文写吗？
谢谢！
j*n
4 楼
07大潮那一拨还有不少人正到了要续EAD, AP的坎上，比如我，O傻再多拖一个月又能进
账不少啊
账不少啊
c*d
5 楼
大概09－10年跳startup的， 有六分之五原地踏步， 六分之一有了一到三米的equity
。 当然还有现在还没ipo的。 回报和跳的时间关系不大，和跳的公司大小关系也不大
， 和水平有一定的关系。 基本上发展好的都是能力情商原来就冒尖的，能力情商高的
也不一定发展的好， 有几个同事去了zynga, groupon就白忙了 。 去linkedin,
facebook的就开心了。跳A B round startup的相对少， fail的可能性也更大。
如果简单的用回报乘以风险，pre-ipo然后股票又在ipo以后涨了是最赚的。 问题是你
不知道上市以后公司股票会怎么走。。。如果你知道不如直接去买股票了。每个人的情
况不一样， 面临的选择也不一样。 这一轮泡泡估计还有一年， 然后象01年， 08年
那样垮下来。我感觉要套现也要抓紧了。
。 当然还有现在还没ipo的。 回报和跳的时间关系不大，和跳的公司大小关系也不大
， 和水平有一定的关系。 基本上发展好的都是能力情商原来就冒尖的，能力情商高的
也不一定发展的好， 有几个同事去了zynga, groupon就白忙了 。 去linkedin,
facebook的就开心了。跳A B round startup的相对少， fail的可能性也更大。
如果简单的用回报乘以风险，pre-ipo然后股票又在ipo以后涨了是最赚的。 问题是你
不知道上市以后公司股票会怎么走。。。如果你知道不如直接去买股票了。每个人的情
况不一样， 面临的选择也不一样。 这一轮泡泡估计还有一年， 然后象01年， 08年
那样垮下来。我感觉要套现也要抓紧了。
j*y
6 楼
我现在的蛋白是个his-tagged homo dimer，我想把 WT 和 active-site mutant 放在
一起组成一个heterodimer.
请问大虾有没有什么好的办法能够得到这样一个homogeneous population 的
heterodimer ?
谢谢
一起组成一个heterodimer.
请问大虾有没有什么好的办法能够得到这样一个homogeneous population 的
heterodimer ?
谢谢
t*s
7 楼
置顶是对的，别处是胡说。
M*s
8 楼
放在schedule A 上。
t*r
9 楼
09年跳回国内阿里巴巴都有3米刀了。人算不如天算啊
s*n
10 楼
理论上可以这样：
构建两个带有不同purification tags而且可以相容的载体，如6xHis tag on pET和
Strep tag on p15A or dual vector。让这两个monomer coexpress，用Ni-NTA和Strep
tag的亲和柱顺序纯化即可。
构建两个带有不同purification tags而且可以相容的载体，如6xHis tag on pET和
Strep tag on p15A or dual vector。让这两个monomer coexpress，用Ni-NTA和Strep
tag的亲和柱顺序纯化即可。
T*a
11 楼
不需要
c*d
12 楼
阿里还没上市， 三米刀怎么算出来的， 按200b valuation?
【在 t**r 的大作中提到】
: 09年跳回国内阿里巴巴都有3米刀了。人算不如天算啊
【在 t**r 的大作中提到】
: 09年跳回国内阿里巴巴都有3米刀了。人算不如天算啊
j*y
13 楼
谢谢楼上的。
我的这个蛋白WT和active-site mutant都是homo-dimer，这样的两个载体co-express能
得到heterodimer吗？会不会只能得两种homodimer？还是两种homodimer加上
heterodimer（minor species)?
我的这个蛋白WT和active-site mutant都是homo-dimer，这样的两个载体co-express能
得到heterodimer吗？会不会只能得两种homodimer？还是两种homodimer加上
heterodimer（minor species)?
h*r
14 楼
谢谢ls两位。
【在 T****a 的大作中提到】
: 不需要
【在 T****a 的大作中提到】
: 不需要
t*r
15 楼
08年入职的很多都有五万股，每股估值66刀，您算算这是多少
【在 c***d 的大作中提到】
: 阿里还没上市， 三米刀怎么算出来的， 按200b valuation?
【在 c***d 的大作中提到】
: 阿里还没上市， 三米刀怎么算出来的， 按200b valuation?
s*n
16 楼
我说的这个方法正是可以解决你的问题，你仔细想一想。
g*g
17 楼
twitter上市前跳都能有 3米刀。
【在 t**r 的大作中提到】
: 09年跳回国内阿里巴巴都有3米刀了。人算不如天算啊
【在 t**r 的大作中提到】
: 09年跳回国内阿里巴巴都有3米刀了。人算不如天算啊
j*y
18 楼
非常感谢，你果然很smart哦:）
c*d
19 楼
扯吧， 不是说三万到五万股的行情。
【在 g*****g 的大作中提到】
: twitter上市前跳都能有 3米刀。
【在 g*****g 的大作中提到】
: twitter上市前跳都能有 3米刀。
a*y
20 楼
you can also use a linker sequence with suitable length to connect one copy
of each gene.
of each gene.
g*g
21 楼
不扯，IPO前半年去能拿到80万 RSU, IPO之后翻了4倍。7万股上下。
【在 c***d 的大作中提到】
: 扯吧， 不是说三万到五万股的行情。
【在 c***d 的大作中提到】
: 扯吧， 不是说三万到五万股的行情。
s*y
22 楼
That is what I would prefer. Some dimer proteins (such as the TdtomatoFP)
was made by this strategy
But smartkevin's method is more simple and straightforward
copy
【在 a**y 的大作中提到】
: you can also use a linker sequence with suitable length to connect one copy
: of each gene.
was made by this strategy
But smartkevin's method is more simple and straightforward
copy
【在 a**y 的大作中提到】
: you can also use a linker sequence with suitable length to connect one copy
: of each gene.
m*a
23 楼
exactly, 不是it泡沫，而是创投泡沫，
这一轮疯狂地VC投资风潮很大一部分是美国的QE和利率政策导致的，有大量的闲置资本
在追逐可怜的一点return。
不过现在看来，利率还会长期低迷，所以创投泡沫破灭的时间可能会比我们预期的晚。
这一轮疯狂地VC投资风潮很大一部分是美国的QE和利率政策导致的，有大量的闲置资本
在追逐可怜的一点return。
不过现在看来，利率还会长期低迷，所以创投泡沫破灭的时间可能会比我们预期的晚。
p*2
24 楼
没有四倍吧
【在 g*****g 的大作中提到】
: 不扯，IPO前半年去能拿到80万 RSU, IPO之后翻了4倍。7万股上下。
【在 g*****g 的大作中提到】
: 不扯，IPO前半年去能拿到80万 RSU, IPO之后翻了4倍。7万股上下。
g*g
25 楼
一度有，现在跌到40多了，但 7万股的话 3M 还是有的。
【在 p*****2 的大作中提到】
: 没有四倍吧
【在 p*****2 的大作中提到】
: 没有四倍吧
p*2
26 楼
一般应该到不了7万股
【在 g*****g 的大作中提到】
: 一度有，现在跌到40多了，但 7万股的话 3M 还是有的。
【在 g*****g 的大作中提到】
: 一度有，现在跌到40多了，但 7万股的话 3M 还是有的。
p*u
27 楼
this is simply impossible for most ppl: just look @ how many fresh grads
Twitter hired before IPO.
【在 g*****g 的大作中提到】
: twitter上市前跳都能有 3米刀。
g*g
28 楼
I am not talking about new grads. I am talking about senior engineers with
10 yrs experience. Regardless, Twitter got 3-4 folds up 6 months after IPO.
That's the kind of pre-ipo companies you want to chase. Low risk, high award
. It can be a hit and miss, and most won't get that kind of return. But you
can always jump after one year. Compared to early startup, the chance to get
IPO or a material acquisition is remote. That's the point I am trying to
make.
【在 p*u 的大作中提到】
:
: this is simply impossible for most ppl: just look @ how many fresh grads
: Twitter hired before IPO.
10 yrs experience. Regardless, Twitter got 3-4 folds up 6 months after IPO.
That's the kind of pre-ipo companies you want to chase. Low risk, high award
. It can be a hit and miss, and most won't get that kind of return. But you
can always jump after one year. Compared to early startup, the chance to get
IPO or a material acquisition is remote. That's the point I am trying to
make.
【在 p*u 的大作中提到】
:
: this is simply impossible for most ppl: just look @ how many fresh grads
: Twitter hired before IPO.
s*a
29 楼
70k RSU package is total bullsxxx, if you are talking about Director
or above position that's a maybe, most common package was 10-15k RSU
for a Sr. level position. Keep in mind IRS & California automatically
took 48% when your RSU vest. A 15k RSU package ends up with $300k after
tax money with 4 years vesting period.
.
award
you
get
【在 g*****g 的大作中提到】
: I am not talking about new grads. I am talking about senior engineers with
: 10 yrs experience. Regardless, Twitter got 3-4 folds up 6 months after IPO.
: That's the kind of pre-ipo companies you want to chase. Low risk, high award
: . It can be a hit and miss, and most won't get that kind of return. But you
: can always jump after one year. Compared to early startup, the chance to get
: IPO or a material acquisition is remote. That's the point I am trying to
: make.
or above position that's a maybe, most common package was 10-15k RSU
for a Sr. level position. Keep in mind IRS & California automatically
took 48% when your RSU vest. A 15k RSU package ends up with $300k after
tax money with 4 years vesting period.
.
award
you
get
【在 g*****g 的大作中提到】
: I am not talking about new grads. I am talking about senior engineers with
: 10 yrs experience. Regardless, Twitter got 3-4 folds up 6 months after IPO.
: That's the kind of pre-ipo companies you want to chase. Low risk, high award
: . It can be a hit and miss, and most won't get that kind of return. But you
: can always jump after one year. Compared to early startup, the chance to get
: IPO or a material acquisition is remote. That's the point I am trying to
: make.
i*i
30 楼
各位大牛指点个明路吧．从目前情况判断，该怎么整？谁家比较有戏？
g*g
31 楼
That's a package preipo, not a package now. $800K RSU over 4 years is not
bullshit. That gives you a package of 400k a year with salary. Good enough
to attract engineers from flg. Airbnb and Uber also give that kind of
package today.
【在 s**a 的大作中提到】
: 70k RSU package is total bullsxxx, if you are talking about Director
: or above position that's a maybe, most common package was 10-15k RSU
: for a Sr. level position. Keep in mind IRS & California automatically
: took 48% when your RSU vest. A 15k RSU package ends up with $300k after
: tax money with 4 years vesting period.
:
: .
: award
: you
: get
bullshit. That gives you a package of 400k a year with salary. Good enough
to attract engineers from flg. Airbnb and Uber also give that kind of
package today.
【在 s**a 的大作中提到】
: 70k RSU package is total bullsxxx, if you are talking about Director
: or above position that's a maybe, most common package was 10-15k RSU
: for a Sr. level position. Keep in mind IRS & California automatically
: took 48% when your RSU vest. A 15k RSU package ends up with $300k after
: tax money with 4 years vesting period.
:
: .
: award
: you
: get
g*g
32 楼
Twitter was worth about $16 in secondary market before it filed IPO, and not
without risk (see FB IPO). If you
think 15K shares or $60K/year will help you compete with FLG for seniors,
think
again.
without risk (see FB IPO). If you
think 15K shares or $60K/year will help you compete with FLG for seniors,
think
again.
s*a
33 楼
Did you get an offer that prints 70k RSU shares on it from Twitter?
If you did not or you never seen one like that, stop imagine numbers here.
not
【在 g*****g 的大作中提到】
: Twitter was worth about $16 in secondary market before it filed IPO, and not
: without risk (see FB IPO). If you
: think 15K shares or $60K/year will help you compete with FLG for seniors,
: think
: again.
If you did not or you never seen one like that, stop imagine numbers here.
not
【在 g*****g 的大作中提到】
: Twitter was worth about $16 in secondary market before it filed IPO, and not
: without risk (see FB IPO). If you
: think 15K shares or $60K/year will help you compete with FLG for seniors,
: think
: again.
s*a
34 楼
70k shares RSU is definitely possible for certain level position (as I said,
Director or principle Engineer level), but no way a regular Sr. SW will get
that.
【在 g*****g 的大作中提到】
: That's a package preipo, not a package now. $800K RSU over 4 years is not
: bullshit. That gives you a package of 400k a year with salary. Good enough
: to attract engineers from flg. Airbnb and Uber also give that kind of
: package today.
Director or principle Engineer level), but no way a regular Sr. SW will get
that.
【在 g*****g 的大作中提到】
: That's a package preipo, not a package now. $800K RSU over 4 years is not
: bullshit. That gives you a package of 400k a year with salary. Good enough
: to attract engineers from flg. Airbnb and Uber also give that kind of
: package today.
f*t
35 楼
T家pre-ipo时entry level就15k，不能说你告诉HR你要think again，他就加到70k的
not
【在 g*****g 的大作中提到】
: Twitter was worth about $16 in secondary market before it filed IPO, and not
: without risk (see FB IPO). If you
: think 15K shares or $60K/year will help you compete with FLG for seniors,
: think
: again.
not
【在 g*****g 的大作中提到】
: Twitter was worth about $16 in secondary market before it filed IPO, and not
: without risk (see FB IPO). If you
: think 15K shares or $60K/year will help you compete with FLG for seniors,
: think
: again.
g*g
36 楼
A friend of mine did, and we had a conversation over it. You don't have to
believe it. I am just telling the truth. People are getting $3M RSU with
Snapchat, and my story of $800K RSU wasn't credible? LOL, you choose what
you want to believe.
http://www.mitbbs.com/article_t1/JobHunting/32724007_0_1.html
【在 s**a 的大作中提到】
: Did you get an offer that prints 70k RSU shares on it from Twitter?
: If you did not or you never seen one like that, stop imagine numbers here.
:
: not
believe it. I am just telling the truth. People are getting $3M RSU with
Snapchat, and my story of $800K RSU wasn't credible? LOL, you choose what
you want to believe.
http://www.mitbbs.com/article_t1/JobHunting/32724007_0_1.html
【在 s**a 的大作中提到】
: Did you get an offer that prints 70k RSU shares on it from Twitter?
: If you did not or you never seen one like that, stop imagine numbers here.
:
: not
g*g
37 楼
像我老混这么久的会跟entry level讨论哪个公司包裹多少钱吗？
【在 f*******t 的大作中提到】
: T家pre-ipo时entry level就15k，不能说你告诉HR你要think again，他就加到70k的
:
: not
【在 f*******t 的大作中提到】
: T家pre-ipo时entry level就15k，不能说你告诉HR你要think again，他就加到70k的
:
: not
s*a
38 楼
你混了这么久了好像还在排期中，我看你也就是个associate等排气的苦主无聊了意淫
一下
【在 g*****g 的大作中提到】
: 像我老混这么久的会跟entry level讨论哪个公司包裹多少钱吗？
一下
【在 g*****g 的大作中提到】
: 像我老混这么久的会跟entry level讨论哪个公司包裹多少钱吗？
s*a
39 楼
that friend then is someone online you heart from...it's something like
"I heard a friend hitting a jackpot of 20 mil on slot machine in Vegas last
month"
【在 g*****g 的大作中提到】
: A friend of mine did, and we had a conversation over it. You don't have to
: believe it. I am just telling the truth. People are getting $3M RSU with
: Snapchat, and my story of $800K RSU wasn't credible? LOL, you choose what
: you want to believe.
: http://www.mitbbs.com/article_t1/JobHunting/32724007_0_1.html
"I heard a friend hitting a jackpot of 20 mil on slot machine in Vegas last
month"
【在 g*****g 的大作中提到】
: A friend of mine did, and we had a conversation over it. You don't have to
: believe it. I am just telling the truth. People are getting $3M RSU with
: Snapchat, and my story of $800K RSU wasn't credible? LOL, you choose what
: you want to believe.
: http://www.mitbbs.com/article_t1/JobHunting/32724007_0_1.html
g*g
40 楼
我老来美国确实不久，才五年，我的同学大多比我早个10年左右。我老说得包裹又不是
什么耸人听闻的东西，要不是 twitter IPO大涨根本没啥好说的。你没见过世面不是我
的错。
【在 s**a 的大作中提到】
: 你混了这么久了好像还在排期中，我看你也就是个associate等排气的苦主无聊了意淫
: 一下
什么耸人听闻的东西，要不是 twitter IPO大涨根本没啥好说的。你没见过世面不是我
的错。
【在 s**a 的大作中提到】
: 你混了这么久了好像还在排期中，我看你也就是个associate等排气的苦主无聊了意淫
: 一下
s*a
41 楼
你来美国前我就在A家了，呵呵。没啥耸人听闻的，就是不太符合行情，我了解的。
【在 g*****g 的大作中提到】
: 我老来美国确实不久，才五年，我的同学大多比我早个10年左右。我老说得包裹又不是
: 什么耸人听闻的东西，要不是 twitter IPO大涨根本没啥好说的。你没见过世面不是我
: 的错。
【在 g*****g 的大作中提到】
: 我老来美国确实不久，才五年，我的同学大多比我早个10年左右。我老说得包裹又不是
: 什么耸人听闻的东西，要不是 twitter IPO大涨根本没啥好说的。你没见过世面不是我
: 的错。
g*g
42 楼
hoho, 我老既没鼓吹自己，又没鼓吹朋友。纯粹说有跟 ali相当的这种机会，就不行了
？ 西雅图才几家公司，你在湾区能让几个 hot start-up互相 bid就有这个行情。
几个进GF早的同学都回国玩风投了。我老至于鼓吹这80万吗？
【在 s**a 的大作中提到】
: 你来美国前我就在A家了，呵呵。没啥耸人听闻的，就是不太符合行情，我了解的。
？ 西雅图才几家公司，你在湾区能让几个 hot start-up互相 bid就有这个行情。
几个进GF早的同学都回国玩风投了。我老至于鼓吹这80万吗？
【在 s**a 的大作中提到】
: 你来美国前我就在A家了，呵呵。没啥耸人听闻的，就是不太符合行情，我了解的。
h*a
43 楼
确实是不错的strategy。
.
award
you
get
【在 g*****g 的大作中提到】
: I am not talking about new grads. I am talking about senior engineers with
: 10 yrs experience. Regardless, Twitter got 3-4 folds up 6 months after IPO.
: That's the kind of pre-ipo companies you want to chase. Low risk, high award
: . It can be a hit and miss, and most won't get that kind of return. But you
: can always jump after one year. Compared to early startup, the chance to get
: IPO or a material acquisition is remote. That's the point I am trying to
: make.
.
award
you
get
【在 g*****g 的大作中提到】
: I am not talking about new grads. I am talking about senior engineers with
: 10 yrs experience. Regardless, Twitter got 3-4 folds up 6 months after IPO.
: That's the kind of pre-ipo companies you want to chase. Low risk, high award
: . It can be a hit and miss, and most won't get that kind of return. But you
: can always jump after one year. Compared to early startup, the chance to get
: IPO or a material acquisition is remote. That's the point I am trying to
: make.
h*a
44 楼
70k shares of RSU was possible in summer 2013. It should be for staff
engineers at least. I know a few cases with similar numbers but certainly
all were pretty senior and had several competing offers.
Interesting thing is that all those engineers I know didn't choose to go to
Twitter. :-)
【在 s**a 的大作中提到】
: 70k RSU package is total bullsxxx, if you are talking about Director
: or above position that's a maybe, most common package was 10-15k RSU
: for a Sr. level position. Keep in mind IRS & California automatically
: took 48% when your RSU vest. A 15k RSU package ends up with $300k after
: tax money with 4 years vesting period.
:
: .
: award
: you
: get
engineers at least. I know a few cases with similar numbers but certainly
all were pretty senior and had several competing offers.
Interesting thing is that all those engineers I know didn't choose to go to
Twitter. :-)
【在 s**a 的大作中提到】
: 70k RSU package is total bullsxxx, if you are talking about Director
: or above position that's a maybe, most common package was 10-15k RSU
: for a Sr. level position. Keep in mind IRS & California automatically
: took 48% when your RSU vest. A 15k RSU package ends up with $300k after
: tax money with 4 years vesting period.
:
: .
: award
: you
: get
w*z
45 楼
你这现身说法啊。
to
【在 h*****a 的大作中提到】
: 70k shares of RSU was possible in summer 2013. It should be for staff
: engineers at least. I know a few cases with similar numbers but certainly
: all were pretty senior and had several competing offers.
: Interesting thing is that all those engineers I know didn't choose to go to
: Twitter. :-)
to
【在 h*****a 的大作中提到】
: 70k shares of RSU was possible in summer 2013. It should be for staff
: engineers at least. I know a few cases with similar numbers but certainly
: all were pretty senior and had several competing offers.
: Interesting thing is that all those engineers I know didn't choose to go to
: Twitter. :-)
h*a
46 楼
ft，我哪有70k啊。不过说实话，如果拿70k offer的同学们能未卜先知Twitter上市后
的走势，估计就接了7万股了。所以发财更多靠运气。
【在 w**z 的大作中提到】
: 你这现身说法啊。
:
: to
的走势，估计就接了7万股了。所以发财更多靠运气。
【在 w**z 的大作中提到】
: 你这现身说法啊。
:
: to
g*g
47 楼
是运气，不过能拿80，只要不跌就不亏。当然像你这样奔退休去的也很好。
【在 h*****a 的大作中提到】
: ft，我哪有70k啊。不过说实话，如果拿70k offer的同学们能未卜先知Twitter上市后
: 的走势，估计就接了7万股了。所以发财更多靠运气。
【在 h*****a 的大作中提到】
: ft，我哪有70k啊。不过说实话，如果拿70k offer的同学们能未卜先知Twitter上市后
: 的走势，估计就接了7万股了。所以发财更多靠运气。
p*2
48 楼
前边有z和g的教训 当时去t的还很怕跌的
涨这么多没人想得到
【在 g*****g 的大作中提到】
: 是运气，不过能拿80，只要不跌就不亏。当然像你这样奔退休去的也很好。
涨这么多没人想得到
【在 g*****g 的大作中提到】
: 是运气，不过能拿80，只要不跌就不亏。当然像你这样奔退休去的也很好。
g*g
49 楼
是呀，我也是马后炮。不过假定不成做一年出来不亏呀。early start up你要是做一年
发现不成基本是白干了。
【在 p*****2 的大作中提到】
: 前边有z和g的教训 当时去t的还很怕跌的
: 涨这么多没人想得到
发现不成基本是白干了。
【在 p*****2 的大作中提到】
: 前边有z和g的教训 当时去t的还很怕跌的
: 涨这么多没人想得到
p*2
50 楼
还是弯曲机会多才能这么搞呀
【在 g*****g 的大作中提到】
: 是呀，我也是马后炮。不过假定不成做一年出来不亏呀。early start up你要是做一年
: 发现不成基本是白干了。
【在 g*****g 的大作中提到】
: 是呀，我也是马后炮。不过假定不成做一年出来不亏呀。early start up你要是做一年
: 发现不成基本是白干了。
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