Newsletter #39

Mike Adams adams at ECSUC.CTSTATEU.EDU
Thu Jan 19 14:20:22 EST 1995

January 1995

This copy of the newsletter is the first that will go via email to the
ChlamyNet. As I indicated in the previous issue all those readers who have
an email account will not be sent a printed version, unless they contact me
and indicate they would like one. Since this is the first issue of the new
year I have also included a copy of the updated mailing list (email readers
can find the list on the BioNet site). Almost all of the items in this
issue have been taken directly from the ChlamyNet and the authors email
address is included in each case.

By now I am sure most of you have heard from Lib Harris concerning the
funding for the Genetics Center. If not, you should be aware that continued
funding by NSF calls for the users (i.e. us) to pay part of the operating
costs. If you use the center Lib asks that you pay a yearly fee of $100.
This can be as a cheque or International Money Order. If you have limited
funding, then Lib asks that you send whatever you can afford.

From: Elizabeth Harris chlamy at acpub.duke.edu
The prototype version of the new Chlamydomonas database is now up and
running on the Plant Genome server at the National Agricultural Library.
There are two ways to reach it:

1)by World Wide Web, URL http://probe.nalusda.gov:8300
2)by gopher to probe.nalusda.gov

There's a pointer on the Chlamy gopher to "Agricultural genome
gopher at NAL" that will take you there also. If you access ChlamyDB via
gopher, select item #2 on probe's main gopher menu ("Plant Genome
Information") and then item #2 again, ("Access to Genome Databases") to
reach the list of available genome databases. All the text information is
contained in both versions.  The WWW version also displays maps of the
nuclear, chloroplast and mitochondrial genomes, and has some additional

Within a few weeks we will also have the database available in a form that
can be transferred by ftp to your own UNIX workstation or high-end
Macintosh.  With this option you will be able to customize the database to
your own use if you like.  You'll find that the displays are nicer this way

From: "Olivier Vallon"  <vallon at ibpc.fr>
 I have found an easy way of getting rid of fungal infections in cultures
of Chlamydomonas reinhardtii. It takes advantage of the fact that
Chlamydomonas  can grow in the presence of methionine sulfoximine (MSX, a
well known inhibitor of glutamine synthase), provided arginine is added to
the medium, while most of our unwanted visitors cannot. If a fungal
infection appears on a plate and it seems difficult to pick a clean
Chlamydomonas  inoculum, streak on a TAP plate supplemented with MSX (100
microM) and arginine (100 mg/l). If possible, place in bright light for 3-4
days, streak back on your usual medium (or a fresh TARG/MSX plate if you
want to be extremely careful). This works fine even with heavily infected
cultures. It seems to work also with bacterial infections. I have no
definite explanation why arginine allows resistance to MSX. In its absence,
50 microM MSX are enough to kill Chlamydomonas  I suspect Chlamydomonas .
has a high ability to use arginine as exclusive ammonium source, bypassing
NH3 and the need for glutamine synthase. Good luck. Let me know of problems
you may encounter, or improvements, or explanations.

From: "Antonio R. Franco" </S=bb1rofra/OU=/@uco.es>
To get rid of bacterial contaminants, I use antibiograms discs. I stripe a
line of Chlamy cells in a TAP solid media and have several of these discs
close of such a line. This way, I can use several antibiotics at the same
time and in the same Petri dish. Tetracycline, Cefotaxime, Trimethoprim,
erythromycin, ceftixozyme are fine. Chlamy cells isolated in this way,
remain in pure cultures for a long time.

From: Julie Nelson julie at molbio.cbs.umn.edu
I have discovered a modification to my transformation protocol using  pJN4,
which is the CRY1-1 gene with a truncated RBCS2 promoter, which  both
improves efficiency and saves time (see Mol. Cell. Biol. 14:  4011-4019,
transformation method 3, for original protocol).  Leave out  the step of
eight hours in SGII right before plating.  So after  transformation, the
steps should be:  3.5 hours in 10 ml SGII, 4 days  in 100 ml SGII-N, pellet
cells, spread onto emetine plates.  The  efficiency improved about two-fold
for me.
From: dutcher at beagle.Colorado.EDU (Susan Dutcher)
The votes have been tabulated and the outcome is below.  I would suggest that
we start to use our new nomenclature rules.

1.  A three-letter designation followed by a number:
        In Favor          69
        Opposed            0

2.  Omitting the hyphen between the acronym (letter) designation and the
    number (abc1)
        In Favor          65
        Opposed            3
        Undecided          1

3.  Pick one alternative:

        Designating locus in upper case letters
        In Favor          48

        Designating locus in lower case letters
        In Favor          20

        Optional           1

4.  Pick one alternative:
        Designating wild-type allele in upper case letters
        In Favor          54

        Designating wild-type allele in lower case letters with a superscript +
        In Favor          13

        Undecided          2
5.  Using d, r, and c to indicate dominant, recessive and codominant alleles,
    if known

        In Favor          55
        Opposed           12
        Undecided          2

6.  Use of the double colon and the inserting DNA to indicate an insertion

        In Favor          54
        Opposed           10
          Optional         5

From: Cliff Zeyl  b7jm at musicb.mcgill.ca
  A few weeks ago I asked for suggestions on why our lab might be having
huge problems cutting genomic DNA from some Chlamy strains with enzymes
that normally aren't too fussy, particularly EcoR1 and Sal1.  Thanks to the
many people who answered.  The most common response was that our DNA
preparations might be contaminated with polysaccharides, apparently a
common problem with preps from plant tissues too.  Among the possible
remedies I tried CTAB precipitation, mostly because it's easy and cheap,
and we do lots of DNA preps.  The method I got was from Current Protocols
in Molecular Biology:  after a phenol-chloroform extraction, add to the
supernatant 1/7 volumes of 5M NaCl, mix VERY well (otherwise the DNA
precipitates), add 0.1 volumes CTAB solution (10% in 0.7M NaCl), and
extract with an equal volume of 24:1 chloroform:isoamyl alcohol.  This has
totally eliminated our problems.

From: rab4m at uva.pcmail.virginia.edu ("Robert A. Bloodgood")

 I have recently become a decision Editor for PROTOPLASMA and wish to take
this opportunity to encourage researchers in the Chlamydomonas community to
consider submitting manuscripts to PROTOPLASMA.  The journal has recently
reorganized its Editorial Board, is expanding its scope and introducing new
and exciting features.  In addition to the regular categories of research
articles, rapid publications and review articles, there is a new feature
called "New Ideas in Cell Biology" which is neither strictly for research
articles or review articles. They are short opinion/discussion pieces on
fast moving and controversial areas of cell biology with no fixed format.
PROTOPLASMA has no page charges, offers one free color plate per article
and provides 100 free reprints per article. You are welcome to make
suggestions for reviewers (and for individuals you would prefer not be used
as reviewers).  Authors are also encouraged to submit a B&W or color
photograph for use on the cover.  Manuscripts for regular research
articles, rapid publication articles, review articles or "New Ideas" pieces
can be sent to me at: Robert A. Bloodgood
Department of Cell Biology
Box 439
University of Virginia Health Sciences Center
Charlottesville, Virginia 22908
Phone:  (804) 924-1739
FAX:    (804) 982-3912
E-mail: RAB4m at Virginia.edu
or to any of the other decision editors listed on the inside cover of each
issue.  If you wish to discuss a possible manuscript or if you wish to have a
copy of the manuscript guidelines sent to you, feel free to contact me by
phone, FAX or E-mail.

Mike Adams
Biology Dept, ECSU
Willimantic, CT 06226
(203) 465-5305

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