Randy Scholl and Keith Davis
Arabidopsis Biological Resource Center, Ohio State University, 309 B&Z Building, 1735 Neil Ave., Columbus, OH 43210, USA
AIMS WWW Server URL: http://aims.cps.msu.edu/aims/
A number of new stocks, released since the publication of the 1996 Supplement to the ABRC catalog, are now available. Descriptions of several of these follow, along with a complete list of the new seed donations. Also, donations of groups of T-DNA lines are needed, and plans for incorporation and use of these are discussed.
ISOLATED DNA FROM FELDMANN T-DNA LINES AVAILABLE
Pools of isolated DNA from the Feldmann T-DNA lines are now being distributed. DNA was isolated from pools of a total of 6,000 T-DNA lines and can be ordered for reverse-genetic PCR screening. The 6,000 lines were divided into six sets of 1000, for which the six pooled DNA samples can be ordered and screened (stock CD5-7). PCR screening of these pools should be conducted with appropriate primer pairs followed by Southern analysis with pBR-free probes, since adjacent insertions are common and the T-DNA contains pBR sequences. For each 1,000 lines, a two-dimensional set of DNAs isolated from pools of 100 plants (i.e, 20 DNA samples) can then be ordered (CD5-11 through CD5-130). Identification of the correct 100-pool in both arrays pinpoints the insertion to a group of ten single lines. The single lines are available as seed samples, from which the investigator can then isolate DNA, conduct PCR to identify the correct insertion line and utilize it for experimentation. Right and left border sequences and the methods employed in the original experiments can be found in McKinney et al. (Plant J. 8:613-622, 1996). A large number of samples of the initial DNAs have already been distributed, and several laboratories have located putative positives. Obviously, many more lines and associated DNAs are needed to improve the likelihood of finding insertions for specific genes. We would be pleased to receive more donations of these resources, as discussed below.
NEW SEED STOCKS RECEIVED
A number of transposon lines have been donated to both ABRC and NASC in recent years. Within the last year, two new sets of Ac/Ds transposon lines with mapped Ds-T-DNA inserts have been received. One of these is from B. Baker, B. Osborne and C. Corr and is described in the 1996 ABRC catalog supplement (available by mail or on-line at http://aims.cps.msu.edu/aims/). The other has been recently received from the Fedoroff laboratory. The map positions of these are linked to the AIMS stock information. It is expected that these lines should be extremely useful for tagging or reverse genetic efforts for cases where the Ds-T-DNA is located near the intended target.
Numbers of new mutant and wild-type lines have been received, recently. These are listed below, along with the current availability status. We are very pleased to acquire these important resources. The majority of these stocks are not yet available, because they are currently being pepared for distribution. You may order the ones which are. Note that this listing is in the order of receipt from April, 1996 to the present. Descriptions of T-DNA lines will be provided later.
Allele/description Donor Availability (stock number) 35S-P1 transgenic B. Krizek available (CS8145) gnom U. Mayer, G. Jürgens available (CS8146) monopterous U. Mayer, G. Jürgens available (CS8147) gurke U. Mayer, G. Jürgens available (CS8148) fackel U. Mayer, G. Jürgens available (CS8149) fass U. Mayer, G. Jürgens available (CS8150) knopf U. Mayer, G. Jürgens available (CS8151) knolle, dis U. Mayer, G. Jürgens available (CS8152) keule U. Mayer, G. Jürgens available (CS8153) stm-1 K. Barton not available rty, rooty J. King not available trp1-1 A. Rose not available trp1-2 A. Rose not available trp1-3 A. Rose not available trp1-5 A. Rose not available trp1-100 A. Rose not available trp1-101 A. Rose not available trp1-102 A. Rose not available trp1-103 A. Rose not available trp1-104 A. Rose not available trp1-105 A. Rose not available trp1-106 A. Rose not available tsl-1 J. Roe not available tsl-2 J. Roe not available mapping lines: TDNA1 M. Van Lijsebettens available (CS5208) TDNA2 M. Van Lijsebettens available (CS5209) TDNA3 M. Van Lijsebettens available (CS5210) TDNA4 M. Van Lijsebettens available (CS5211) TDNA5 M. Van Lijsebettens available (CS5212) TDNA6 M. Van Lijsebettens available (CS5213) TDNA7 M. Van Lijsebettens available (CS5214) TDNA8 M. Van Lijsebettens available (CS5215) ecotype lines sampled from several locations R. Mauricio not available amp1-1 A. Chin-Atkins not available uvr2 R. Last available (CS8525) soz1 R. Last available (CS8526) trp2-1 R. Last not available trp2-8 R. Last not available trp2-11 R. Last not available trp2-100 R. Last not available trp3-1 R. Last not available trp3-100 R. Last not available 180 transposed Ds lines D. Söll not available mapped Ds transposon and Ac transposase lines N. Fedoroff available (CS8510-CS8538) scr-1 P. Benfey not available scr-2 P. Benfey not available cob-1 P. Benfey not available cob-2 P. Benfey not available cob-3 P. Benfey not available AEQ transgenic J. Braam not available rss 3B J. Werner-Fracjek not available rss 5A J. Werner-Fracjek not available rss 6A J. Werner-Fracjek not available rss 7A J. Werner-Fracjek not available rss 9A J. Werner-Fracjek not available rss 11A J. Werner-Fracjek not available ett-1 A. Sessions not available ett-2 A. Sessions not available ett-3 A. Sessions not available
The stocks on this list which are currently not available will be released as soon as possible. This may take up to several months for stocks which are maintained as heterozygotes. In all cases notification of availability will be made via AIMS and the newsgroup.
NEW DNA STOCKS NOW AVAILABLE
Numbers of new clones have also been received. The new DNA stocks which are now available are described below.
1. Mapping clones donated by Mitsui Plant Biotechnology Research Institute. (YG. Liu, N. Mitsukawa, C. Lister, C. Dean and R. F. Whittier. 1996. Plant J. 10:733735)
This large new collection of plasmid mapping clones is now available for shipping. These should be especially useful for individuals interested in positional cloning. Consult the DNA search page http://aims.cps.msu.edu/aims/menu/search2.html to order these clones and obtain information.
Stock # clone name Map position CD3-205 mi15 1-55.7 CD3-206 mi19 1-75.9 CD3-207 mi30 4-73.1 CD3-208 mi32 4-84.5 CD3-209 mi51 4-11 CD3-210 mi54 2-55.1 CD3-211 mi61 5-101.1 CD3--212 mi62 1-55.7 CD3--213 mi63 1-74.9 CD3-214 mi69 5-122.1 CD3-215 mi70 5-123.2 CD3-216 mi72 1-81.7 CD3-217 mi74 3-1.2 CD3-218 mi79a 2-82 CD3-219 mi79b 3-70.3 CD3-220 mi83 5-97.2 CD3-221 mi87 4-42.5 CD3-222 mi90 5-36.7 CD3-223 mi97 5-13 CD3-224 mi100 1-16 CD3-225 mi103 1-137.5 CD3-226 mi106 1-98.2 CD3-227 mi107 1-55.7 CD3-228 mi111 1-55.7 CD3-229 mi112 4-81.8 CD3-230 mi116 1-56.7 CD3-231 mi121 5-8.0 CD3-232 mi122 4-14.5 CD3-233 mi123 4-100.9 CD3-234 mi125 5-60.3 CD3-235 mi128 4-69.5 CD3-236 mi133 1-79.3 CD3-237 mi137 5-69.9 CD3-238 mi138 5-32.4 CD3-239 mi139 2-34.3 CD3-240 mi142 3-38.6 CD3-241 mi148 2-35.4 CD3-242 mi157 1-152.6 CD3-243 mi163 1-52 CD3-244 mi167 4-42 CD3-245 mi172 3-3.8 CD3-246 mi174 5-21 CD3-247 mi178 3-59.6 CD3-248 mi184 5-123.2 CD3-249 mi185 1-125.8 CD3-250 mi192 1-55.7 CD3-251 mi193 1-125.8 CD3-252 mi194 5-96.2 CD3-253 mi198 4-76.1 CD3-254 mi199 3-2.2 CD3-255 mi203 1-35 CD3-256 mi204 4-13.9 CD3-257 mi207 3-18.9 CD3-258 mi208 1-97.7 CD3-260 mi219 5-44.6 CD3-261 mi225 3-48.8 CD3-262 mi230 1-109.2 CD3-263 mi232 4-102 CD3-264 mi233 4-38.4 CD3-265 mi238 2-38.1 CD3-266 mi259 1-109.2 CD3-267 mi260 4-78.2 CD3-268 mi265 1-51.4 CD3-269 mi268 3-40.3 CD3-270 mi277 2-65.7 CD3-271 mi279 4-72.6 CD3-272 mi287 3-59.6 CD3-273 mi289 3-25.4 CD3-274 mi291a 1-96.1 CD3-275 mi291b 5-67.2 CD3-276 mi301 4-23.9 CD3-277 mi303 1-106.7 CD3-278 mi304 1-109.2 CD3-279 mi306 4-39.9 CD3-280 mi310 2-17.9 CD3-281 mi320 2-9.3 CD3-282 mi322 5-26.9 CD3-283 mi323 5-79.7 CD3-284 mi324 1-114.5 CD3-285 mi328 2-9.3 CD3-286 mi330 4-81.8 CD3-287 mi334 4-39.9 CD3-288 mi335 5-143.6 CD3-289 mi339 3-25.9 CD3-290 mi342 1-76.9 CD3-291 mi348 1-30.8 CD3-292 mi353 1-114.5 CD3-293 mi355 3-9.6 CD3-294 mi357 3-10.7 CD3-295 mi358 3-69.8 CD3-296 mi369 4-139.3 CD3-297 mi372 1-10.6 CD3-298 mi386 3-47.6 CD3-299 mi390 4-25.9 CD3-300 mi398 2-28.5 CD3-301 mi403 3-9.6 CD3-302 mi408 1-109.2 CD3-303 mi413 3-67.2 CD3-304 mi418 5-123.2 CD3-305 mi421 2-18.5 CD3-306 mi422 4-92.8 CD3-307 mi423a 1-71.6 CD3-308 mi423b 5-100.5 CD3-309 mi424 1-116.2 CD3-310 mi425 1-143.4 CD3-311 mi431 4-107.5 CD3-312 mi433 5-36.2 CD3-313 mi438 5-26.9 CD3-314 mi441 1-97.2 CD3-315 mi443 1-16.6 CD3-316 mi444 2-18.5 CD3-317 mi455 2-82 CD3-318 mi456 3-101.4 CD3-319 mi462 1-136.5 CD3-320 mi465 4-66.7 CD3-321 mi467 3-13.7 CD3-322 mi473 2-83.6
2. Green fluorescent protein clones from the R. Vierstra laboratory (S. J. Davis and R. D. Vierstra. Weeds World vol. 3ii.)
These useful reporter constructs are now available for distribution. The original protein was modified for solubilization as well as shift of the spectrum of the fluorescence. The ordering location for these stocks is http://aims.cps.msu.edu/aims/menu/search2.html. The three stocks are:
CD3-326 solublemodified GFP CD3-327 - solublemodifed and red shifted GFP CD3-328 solublemodified Blue FP
T-DNA LINE ACQUISITIONS AND PLANS
The stock centers are currently striving to increase holdings of T-DNA lines. In this regard, ABRC is currently processing 2,000+ new T-DNA lines provided by Thomas Jack, which should be ready for distribution around April, 1997. An announcement will be made on the Arabidopsis newsgroup when these are available. We are also receiving some lines from other laboratories and hope to substantially increase the number of available T-DNA lines and derived DNAs during the next year. We invite all individuals who have T-DNA populations which can be shared to contact us, and we will cooperate to make these available to all researchers. We need both pools of lines for phenotypic screening and DNA for reverse genetics. Please let us know if you have any such material which you can share.
Our current plan for handling T-DNA donations is as follows. We will pool the seeds of individual lines received into pools of ten and grow a large stock from these. Resulting pools of ten will be combined into groups of 100 for initial phenotypic screening. Hence, initial phenotypic screening will be conducted at this level with associated 10-pools being available for rescreening. In cases where sufficient seeds of individual lines are donated, the remnant seed will constitute a stock for followup single-line evaluations. DNA will be isolated from the same 100-pools used for phenotypic screening as well as a second dimension within sets of 1000 lines. This will allow location of an insertion to a 10-pool with minimal PCR analysis.
The general scheme described above is designed for donations of large (1000+) sets of individual lines. It would also be very useful for donations of lines already pooled into sets of size 10 or 20. In the latter cases, the lowest level of screening available to the investigator would be a pool of 10 or 20 lines. Based on this general model, we believe that we can process several thousand lines per year for both phenotypic and PCR screening. Donations of lines for this purpose are needed immediately, and we especially would like to receive seeds in large enough quantities for immediate distribution as well as already-isolated DNA if these can be spared!
As always, new donations of all seed and DNA stocks are welcome. The publicly available collections are now growing rapidly, but there are many mutants and clones which we would very much like to receive.